Elaine L Wilson, Ph.D.

Molecular signatures of the primitive prostate stem cell niche reveal novel mesenchymal-epithelial signaling pathways. L
Blum, Roy; Gupta, Rashmi; Burger, Patricia E; Ontiveros, Christopher S; Salm, Sarah N; Xiong, Xiaozhong; Kamb, Alexander; Wesche, Holger; Marshall, Lisa; Cutler, Gene; Wang, Xiangyun; Zavadil, Jiri; Moscatelli, David; Wilson, E Lynette
2010 ;5(9):?-?, PLoS ONE
BACKGROUND: Signals between stem cells and stroma are important in establishing the stem cell niche. However, very little is known about the regulation of any mammalian stem cell niche as pure isolates of stem cells and their adjacent mesenchyme are not readily available. The prostate offers a unique model to study signals between stem cells and their adjacent stroma as in the embryonic prostate stem cell niche, the urogenital sinus mesenchyme is easily separated from the epithelial stem cells. Here we investigate the distinctive molecular signals of these two stem cell compartments in a mammalian system. METHODOLOGY/PRINCIPAL FINDINGS: We isolated fetal murine urogenital sinus epithelium and urogenital sinus mesenchyme and determined their differentially expressed genes. To distinguish transcripts that are shared by other developing epithelial/mesenchymal compartments from those that pertain to the prostate stem cell niche, we also determined the global gene expression of epidermis and dermis of the same embryos. Our analysis indicates that several of the key transcriptional components that are predicted to be active in the embryonic prostate stem cell niche regulate processes such as self-renewal (e.g., E2f and Ap2), lipid metabolism (e.g., Srebp1) and cell migration (e.g., Areb6 and Rreb1). Several of the enriched promoter binding motifs are shared between the prostate epithelial/mesenchymal compartments and their epidermis/dermis counterparts, indicating their likely relevance in epithelial/mesenchymal signaling in primitive cellular compartments. Based on differential gene expression we also defined ligand-receptor interactions that may be part of the molecular interplay of the embryonic prostate stem cell niche. CONCLUSIONS/SIGNIFICANCE: We provide a comprehensive description of the transcriptional program of the major regulators that are likely to control the cellular interactions in the embryonic prostatic stem cell niche, many of which may be common to mammalian niches in general. This study provides a comprehensive source for further studies of mesenchymal/epithelial interactions in the prostate stem cell niche. The elucidation of pathways in the normal primitive niche may provide greater insight into mechanisms subverted during abnormal proliferative and oncogenic processes. Understanding these events may result in the development of specific targeted therapies for prostatic diseases such as benign prostatic hypertrophy and carcinomas
— id: 113814, year: 2010, vol: 5, page: ?, stat: Journal Article,

PINing Down the Origin of Prostate Cancer
Moscatelli, David; Wilson, E Lynette
2010 Aug 4;2(43):43ps38-43ps38, Science translational medicine
The epithelium that lines the surface of prostate glands contains several cell types, including luminal secretory cells and basal cells of unclear function. Despite the fact that prostate tumors contain cells with a luminal phenotype and lack basal cells, a recent report indicates that the cell of origin for human prostate cancer is a basal cell and not a luminal cell. In contrast, another study indicates the reverse. It is possible that both basal and luminal stem/progenitor cells may independently give rise to prostate cancer; a comparison of the molecular signatures of the target cells of transformation with those of prostate tumors may aid in predicting the phenotypes of tumors with aggressive characteristics
— id: 111550, year: 2010, vol: 2, page: 43ps38, stat: Journal Article,

Molecular signatures of prostate stem cells reveal novel signaling pathways and provide insights into prostate cancer
Blum, Roy; Gupta, Rashmi; Burger, Patricia E; Ontiveros, Christopher S; Salm, Sarah N; Xiong, Xiaozhong; Kamb, Alexander; Wesche, Holger; Marshall, Lisa; Cutler, Gene; Wang, Xiangyun; Zavadil, Jiri; Moscatelli, David; Wilson, E Lynette
2009 ;4(5):e5722-e5722, PLoS ONE
BACKGROUND: The global gene expression profiles of adult and fetal murine prostate stem cells were determined to define common and unique regulators whose misexpression might play a role in the development of prostate cancer. METHODOLOGY/PRINCIPAL FINDINGS: A distinctive core of transcriptional regulators common to both fetal and adult primitive prostate cells was identified as well as molecules that are exclusive to each population. Elements common to fetal and adult prostate stem cells include expression profiles of Wnt, Shh and other pathways identified in stem cells of other organs, signatures of the aryl-hydrocarbon receptor, and up-regulation of components of the aldehyde dehydrogenase/retinoic acid receptor axis. There is also a significant lipid metabolism signature, marked by overexpression of lipid metabolizing enzymes and the presence of the binding motif for Srebp1. The fetal stem cell population, characterized by more rapid proliferation and self-renewal, expresses regulators of the cell cycle, such as E2f, Nfy, Tead2 and Ap2, at elevated levels, while adult stem cells show a signature in which TGF-beta has a prominent role. Finally, comparison of the signatures of primitive prostate cells with previously described profiles of human prostate tumors identified stem cell molecules and pathways with deregulated expression in prostate tumors including chromatin modifiers and the oncogene, Erg. CONCLUSIONS/SIGNIFICANCE: Our data indicate that adult prostate stem or progenitor cells may acquire characteristics of self-renewing primitive fetal prostate cells during oncogenesis and suggest that aberrant activation of components of prostate stem cell pathways may contribute to the development of prostate tumors
— id: 99241, year: 2009, vol: 4, page: e5722, stat: Journal Article,

High Aldehyde Dehydrogenase Activity: A Novel Functional Marker of Murine Prostate Stem/Progenitor Cells
Burger, PE; Gupta, R; Xiong, X; Ontiveros, CS; Salm, SN; Moscatelli, D; Wilson, EL
2009 OCT ;27(9):2220-2228, Stem cells
We have shown previously that prostatic stem/progenitor cells can be purified from isolated prostate ducts, based on their high expression of the Sca-1 surface antigen. We now report that high levels of aldehyde dehydrogenase (ALDH) activity are present in a subset of prostate epithelial cells that coexpress a number of antigens found on stem/progenitor cells of other origins (CD9, Bcl-2, CD200, CD24, prominin, Oct 3/4, ABCG2, and nestin). Almost all of these cells expressing high levels of ALDH activity also express Sca-1 and a third of them express high levels of this antigen. The cells with high levels of ALDH activity have greater in vitro proliferative potential than cells with low ALDH activity. Importantly, in an in vivo prostate reconstitution assay, the cells expressing high levels of ALDH activity were much more effective in generating prostatic tissue than a population of cells with low enzymatic activity. Thus, a high level of ALDH activity can be considered a functional marker of prostate stem/progenitor cells and allows for simple, efficient isolation of cells with primitive features. The elucidation of the role of ALDH in prostate stem/progenitor cells may lead to the development of rational therapies for treating prostate cancer and benign prostatic hyperplasia. STEM CELLS 2009; 27: 2220-2228
— id: 102959, year: 2009, vol: 27, page: 2220, stat: Journal Article,

Endothelial cells support the growth of prostate tissue in vivo
Bates, Michael; Kovalenko, Bruce; Wilson, E Lynette; Moscatelli, David
2008 Jun 1;68(8):893-901, Prostate
INTRODUCTION: The contribution of vascular endothelial cells to prostate growth has not been investigated. We examined whether endothelial cells support growth of prostate tissue when co-inoculated with prostate epithelial cells under the renal capsule. METHODS: Vascular endothelial cells were isolated from mice and co-inoculated under the renal capsule with a prostate luminal or basal epithelial cell line. After 60 days, kidneys were examined for growth of prostate tissue. Prostatic tissues were examined by immunohistochemistry for expression of cytokeratins 5 and 8, and vascular density was determined. To determine if increased expression of VEGF-A would increase prostatic growth, transfected endothelial cells overexpressing VEGF-A were co-inoculated with the prostate luminal or basal epithelial lines. RESULTS: Co-inoculation of endothelial cells and prostate luminal or basal epithelial cells resulted in significant growth of prostatic tissue, whereas inoculation of any of the cell lines alone resulted in little growth. The growths from co-inoculation of endothelial cells and luminal epithelial cells contained duct-like structures that stained with antibodies to cytokeratin 8, whereas those from co-inoculation of endothelial cells and basal epithelial cells contained cords of cells that stained with antibodies to cytokeratin 5. Overexpression of VEGF-A had no effect on growth of the prostatic tissues. CONCLUSION: Endothelial cells contribute to the growth of prostatic epithelial cells
— id: 79381, year: 2008, vol: 68, page: 893, stat: Journal Article,

Axin2 expression identifies progenitor cells in the murine prostate
Ontiveros, Christopher S; Salm, Sarah N; Wilson, E Lynette
2008 Sep 1;68(12):1263-1272, Prostate
BACKGROUND: We previously reported that prostatic stem/progenitor cells are concentrated in the proximal region of prostatic ducts and express stem cell antigen 1 (Sca-1). As Wnt signaling is important for the maintenance of stem cells, we determined whether Sca-1 expressing cells also express Axin2, as Axin2 expression is highly suggestive of active Wnt signaling. METHODS: Axin2 promoter reporter mice were used for whole mount and fluorescence activated cell sorting (FACS) analysis to determine its expression in the prostate. Axin2 expressing cells were also examined for the co-expression of Sca-1. We also used a chemical activator of Wnt signaling, BIO, to determine the effects of Wnt signaling on the growth of primary prostate cells in vitro. RESULTS: We show that Axin2 expression is present in all lobes and is regulated by androgens with the highest Axin2 expression in the lateral and dorsal prostate. Furthermore, a fraction of Axin2 expressing cells co-express Sca-1, suggesting that some progenitor cells have active Wnt signaling. Lastly, we demonstrate that activation of the Wnt pathway may result in increased growth, consistent with a role for Wnt signaling in maintenance and/or expansion of the progenitor cell population. CONCLUSION: Axin2 expressing cells that co-express Sca-1 are present in all prostate lobes suggesting that progenitor cells reside within the Wnt active population. An understanding of the basic biology of signaling pathways mediating growth in the prostate may lead to rational therapies to treat benign prostatic hyperplasia and prostate cancer
— id: 84018, year: 2008, vol: 68, page: 1263, stat: Journal Article,

Vascular density is highest in the proximal region of the mouse prostate
Wang, Gui-Min; Kovalenko, Bruce; Wilson, E Lynette; Moscatelli, David
2007 Jun 15;67(9):968-975, Prostate
BACKGROUND: The proximal region of the prostatic ducts harbor the prostatic epithelial stem cells. As stem cell niches in other organs are highly vascularized, we determined if the proximal region is more highly vascularized than the remaining regions of the prostate. The effect of androgen on vascular density in the different prostatic regions was also examined. METHODS: Sections from prostates were immunostained with antibodies to CD31, and the vascular density in proximal, intermediate, and distal regions was calculated by image analysis software. Vascular density was compared in prostates from castrated mice that received daily inoculations of testosterone or vehicle alone for 3 days. To examine the role of angiogenic factors in the response to androgen, some animals were also treated with soluble VEGF receptor-2-Fc or Tie-2--Fc fusion proteins, which inhibit the activities of VEGF and angiopoietins, respectively. The endothelial proliferative response to androgen was determined by double staining sections with antibodies to CD31 and Ki-67. RESULTS: In prostates from intact mice, vascular density was highest in the proximal region and lowest in the distal region. Administration of testosterone to castrated mice increased vascular density to the greatest extent in the distal and intermediate regions. The increase in vascular density required VEGF and the angiopoietins. Endothelial cell proliferation was less sensitive to androgen in the proximal region than the remainder of the prostate. CONCLUSIONS: Vascular density is highest in the proximal region of the prostate, but the proximal vessels are less responsive to testosterone.
— id: 72722, year: 2007, vol: 67, page: 968, stat: Journal Article,

Proximal prostatic stem cells are programmed to regenerate a proximal-distal ductal axis
Goto, Ken; Salm, Sarah N; Coetzee, Sandra; Xiong, Xiaozhong; Burger, Patricia E; Shapiro, Ellen; Lepor, Herbert; Moscatelli, David; Wilson, E Lynette
2006 Aug;24(8):1859-1868, Stem cells
Prostate carcinoma and benign prostatic hypertrophy may both originate in stem cells, highlighting the importance of the characterization of these cells. The prostate gland contains a network of ducts each of which consists of a proximal (adjacent to the urethra), an intermediate, and a distal region. Here, we report that two populations of cells capable of regenerating prostatic tissue in an in vivo prostate reconstitution assay are present in different regions of prostatic ducts. The first population (with considerable growth potential) resides in the proximal region of ducts and in the urethra, and the survival of these cells does not require the presence of androgens. The second population (with more limited growth potential) is found in the remaining ductal regions and requires androgen for survival. In addition, we find that primitive proximal prostate cells that are able to regenerate functional prostatic tissue in vivo are also programmed to re-establish a proximal-distal ductal axis. Similar to their localization in the intact prostate, cells with the highest regenerative capacity are found in the proximal region of prostatic ducts formed in an in vivo prostate reconstitution assay. The primitive proximal cells can be passaged through four generations of subrenal capsule grafts. Together, these novel findings illustrate features of primitive prostate cells that may have implications for the development of therapies for treating proliferative prostatic diseases
— id: 72058, year: 2006, vol: 24, page: 1859, stat: Journal Article,

Sca-1 expression identifies stem cells in the proximal region of prostatic ducts with high capacity to reconstitute prostatic tissue
Burger, Patricia E; Xiong, Xiaozhong; Coetzee, Sandra; Salm, Sarah N; Moscatelli, David; Goto, Ken; Wilson, E Lynette
2005 May 17;102(20):7180-7185, Proceedings of the National Academy of Sciences of the United States of America
We previously showed that prostatic stem cells are concentrated in the proximal regions of prostatic ducts. We now report that these stem cells can be purified from isolated proximal duct regions by virtue of their high expression of the cell surface protein stem cell antigen 1 (Sca-1). In an in vivo prostate reconstitution assay, the purified Sca-1-expressing cell population isolated from the proximal region of ducts was more effective in generating prostatic tissue than a comparable population of Sca-1-depleted cells (203.0 +/- 83.1 mg vs. 11.9 +/- 9.2 mg) or a population of Sca-1-expressing cells isolated from the remaining regions of ducts (transit-amplifying cells) (31.9 +/- 24.1 mg). Almost all of the proliferative capacity of the proximal duct Sca-1-expressing cell population resides within the fraction of cells that express high levels of Sca-1 (top one-third), with the proximal region of prostatic ducts containing 7.2-fold more Sca-1(high) cells than the remaining regions. More than 60% of the high-expressing cells coexpress alpha6 integrin and the anti-apoptotic factor Bcl-2, markers that are also characteristic of stem cells of other origins. Further stratification of the phenotype of the stem cells may enable the development of rational therapies for treating prostate cancer and benign prostatic hyperplasia.
— id: 72723, year: 2005, vol: 102, page: 7180, stat: Journal Article,

TGF-{beta} maintains dormancy of prostatic stem cells in the proximal region of ducts
Salm, Sarah N; Burger, Patricia E; Coetzee, Sandra; Goto, Ken; Moscatelli, David; Wilson, E Lynette
2005 Jul;170(1):81-90, Journal of cell biology
We have previously shown that prostatic stem cells are located in the proximal region of mouse prostatic ducts. Here, we show that this region responds differently to transforming growth factor (TGF)-beta than the distal ductal region and that under physiological conditions androgens and TGF-beta are crucial overall regulators of prostatic tissue homeostasis. This conclusion is supported by the observations showing that high levels of TGF-beta signaling are present in the quiescent proximal region of ducts in an androgen-replete animal and that cells in this region overexpress Bcl-2, which protects them from apoptosis. Moreover, androgen ablation reverses the proximal-distal TGF-beta signaling gradient, leading to an increase in TGF-beta signaling in the unprotected distal region (low Bcl-2 expression). This reversal of TGF-beta-mediated signaling accompanies apoptosis of cells in the distal region and gland involution after androgen withdrawal. A physiological TGF-beta signaling gradient (high proximally and low distally) and its functional correlates are restored after androgen replenishment. In addition to highlighting the regulatory role of androgens and TGF-beta, these findings may have important implications for the deregulation of the stem cell compartment in the etiology of proliferative prostatic diseases
— id: 56205, year: 2005, vol: 170, page: 81, stat: Journal Article,

IMMUNOLOCALIZATION OF ESTROGEN RECEPTOR alpha AND beta IN HUMAN FETAL PROSTATE
Shapiro, Ellen; Huang, Hongying; Masch, Rachel J; McFadden, Deborah E; Wilson, E Lynette; Wu, Xue-Ru
2005 Nov;174(5):2051-2053, Journal of urology
PURPOSE:: We examined the immunolocalization of estrogen receptor (ER)alpha and ERbeta in the human fetal prostate. MATERIALS AND METHODS:: Tissue sections from human fetal prostates at 7 to 22 weeks of gestation were stained with antibodies to ERalpha, ERbeta, and cytokeratin 10 and 14. RESULTS:: ERalpha expression was not detected until 15 weeks of gestation with sparse staining in the utricle. By 19 weeks increased ERalpha expression was seen in the luminal cells of the ventral urogenital epithelium (UGE), basal cells of the dorsal UGE, utricle, distal periurethral ducts, peripheral stroma and posterior prostatic duct. K14 was detected in basal cells of the UGE and in several posterior acini. At 22 weeks ERalpha expression was more intense in all of these areas. ERbeta was expressed throughout the UGE, ejaculatory ducts, mullerian ducts and entire stroma at 7 weeks. Intense ERbeta staining was observed in these areas and in the prostatic buds by 8 weeks with persistent intense staining through 22 weeks. CONCLUSIONS:: To our knowledge we report the first immunolocalization of ERalpha in the human fetal prostate and the earliest demonstration of ERbeta expression in the prostate at 7 weeks of gestation. ERbeta expression is intense during ductal morphogenesis, suggesting a role in normal glandular growth and proliferation. The induction of squamous metaplasia in the UGE, distal periurethral ducts and utricle is associated with ERalpha expression in these areas, while the induction of squamous metaplasia in peripheral prostatic acini is associated with peripheral stromal ERalpha expression. This study suggests estrogen signaling pathways in the human fetal prostate via ERalpha that involve epithelial-epithelial and epithelial-stromal interactions
— id: 58459, year: 2005, vol: 174, page: 2051, stat: Journal Article,

Stromal/epithelial interactions of murine prostatic cell lines in vivo: a model for benign prostatic hyperplasia and the effect of doxazosin on tissue size
Takao, Tetsuya; Tsujimura, Akira; Coetzee, Sandra; Salm, Sarah N; Lepor, Herbert; Shapiro, Ellen; Moscatelli, David; Wilson, E Lynette
2003 Jan 1;54(1):17-24, Prostate
BACKGROUND: One of the major constraints in elucidating the mechanisms involved in the etiology of benign prostatic hyperplasia (BPH) is the lack of suitable model systems that are readily manipulable in vitro and in vivo. To address this issue, we have used murine prostatic cell lines to establish a novel in vivo model for studying prostatic cell interactions. METHODS: Luminal, basal, and smooth muscle (SM) cell lines were inoculated alone or in combinations under the renal capsule of intact or castrated male mice, and the growth and composition of prostatic tissue in the absence or presence of doxazosin was determined. RESULTS: Both the luminal and basal cell lines reconstituted prostatic tissue if co-inoculated under the renal capsule with normal SM cells, whereas none of the lines formed significant tissue when inoculated alone. Luminal cells produced and secreted prostatic secretory products. The growth of prostatic tissue formed from co-inoculation of basal and SM cells was androgen responsive. In addition, a significant reduction in prostatic tissue was noted in animals treated with doxazosin. CONCLUSION: We have established an in vivo model that uses prostatic epithelial and SM cell lines for investigating cellular interactions between epithelial and SM cells that regulate prostatic growth and function. This model will be useful for delineating the mechanisms by which prostatic cells interact and in determining the efficacy of new approaches aimed at interfering with prostatic stromal/epithelial interactions that result in abnormal cellular proliferation
— id: 35189, year: 2003, vol: 54, page: 17, stat: Journal Article,

Fibroblast growth factor receptor-1 is expressed by endothelial progenitor cells
Burger, Patricia E; Coetzee, Sandra; McKeehan, Wallace L; Kan, Mikio; Cook, Perry; Fan, Yong; Suda, Toshio; Hebbel, Robert P; Novitzky, Nicolas; Muller, William A; Wilson, E Lynette
2002 Nov 15;100(10):3527-3535, Blood
Recent experiments show that hematopoietic progenitor cell populations contain endothelial precursor cells. We have isolated a population of CD34(+) cells that expresses fibroblast growth factor receptor-1 (FGFR-1) and that differentiates into endothelial cells in vitro. We find that 4.5% +/- 2.1% of CD34(+) cells isolated from bone marrow, cord blood, and mobilized peripheral blood express FGFR-1 and that viable CD34(+)FGFR(+) cells are small, with little granularity, and express both primitive hematopoietic and endothelial markers on their surface. The primitive hematopoietic markers AC133, c-kit, and Thy-1 are coexpressed by 75%, 85%, and 64% of CD34(+)FGFR(+) cells, respectively. Most of the CD34(+)FGFR(+) cells also express antigens found on endothelial cells, such as CD31, vascular endothelial growth factor receptor-2, and the endothelial-specific cell surface marker, vascular endothelial cadherin (VE-cadherin), whereas 56% to 60% of the cells express Tie, Tek, and the endothelial-specific marker, P1H12. The CD34(+)FGFR(+) population is enriched in cells expressing endothelial-specific antigens compared with the CD34(+) population. Isolated CD34(+)FGFR(+) cells grow slowly in culture, are stimulated by fibroblast growth factor-2 and vascular endothelial growth factor, and give rise to cells that express von Willebrand factor and VE-cadherin and that incorporate acetylated low-density lipoprotein. These experiments show that FGFR-1 is expressed by a subpopulation of CD34(+) cells that give rise to endothelial cells in vitro, indicating that this population contains endothelial stem/progenitor cells
— id: 35190, year: 2002, vol: 100, page: 3527, stat: Journal Article,

Basic fibroblast growth factor modulates the expression of glycophorin A and c-kit and inhibits erythroid differentiation in K562 cells
Burger, Patricia E; Lukey, Pauline T; Coetzee, Sandra; Wilson, E Lynette
2002 Jan;190(1):83-91, Journal of cellular physiology
Basic fibroblast growth factor (bFGF) is produced by bone marrow stromal cells as well as by normal and leukemic hematopoietic cells. In this study, we examine the direct effects of bFGF on erythroid differentiation in K562 cells in order to determine whether bFGF can promote the expression of a primitive phenotype. Low levels of bFGF inhibited erythroid differentiation as evidenced by decreased expression of glycophorin A and increased expression of c-kit. bFGF also increased both the numbers and the sizes of colonies of K562 cells in soft agar assays. The addition of TGF-beta to these cells induced erythroid differentiation which resulted in an increase in glycophorin A and a decrease in c-kit. The simultaneous addition of bFGF and TGF-beta to K562 cells prevented both the TGF-beta-mediated increase in glycophorin A expression and the decrease in c-kit expression associated with erythroid differentiation. bFGF antagonised the TGF-beta-mediated promotion of erythroid differentiation in K562 cells in a dose dependent manner and these two cytokines counteracted each other on an approximately molar basis. These results indicate that bFGF alone increases expression of c-kit and promotes a primitive phenotype in K562 cells. In addition, bFGF counteracts the effects of differentiation-inducing cytokines, such as TGF-beta, on hematopoietic cells. It is therefore possible that enhanced production of bFGF by leukemic cells could contribute to their neoplastic phenotype by opposing the effects of negative regulators or cytokines that induce differentiation
— id: 35193, year: 2002, vol: 190, page: 83, stat: Journal Article,

Androgens modulate the balance between VEGF and angiopoietin expression in prostate epithelial and smooth muscle cells
Richard, Christian; Kim, Gilbert; Koikawa, Yasuhiro; Salm, Sarah N; Tsujimura, Akira; Wilson, E Lynette; Moscatelli, David
2002 Feb 1;50(2):83-91, Prostate
BACKGROUND: The vasculature of the prostate responds to androgens. Androgens most likely affect the vasculature indirectly by modulating the expression of angiogenic factors in the cells of the prostate. Most studies to date have examined the production of angiogenic factors by the prostate luminal epithelium. Here we examine the effects of androgen on production of three angiogenic factors, vascular endothelial growth factor (VEGF), angiopoietin-1, and angiopoietin-2, by the three major cell types in the prostate. METHODS: The ability of androgen to modulate VEGF, angiopoietin-1, and angiopoietin-2 production in cultured mouse prostate luminal epithelial, basal epithelial, and smooth muscle cells (SMCs) was assessed by Western blot and RT-PCR. RESULTS: The production of VEGF was modulated by androgens in both luminal epithelial and prostate SMCs but not in basal epithelial cells. However, in prostate luminal epithelial cell cultures, VEGF was predominately secreted apically, suggesting that in vivo most of the epithelium-derived VEGF is unavailable to the underlying blood vessels. In addition, prostate luminal epithelial cells produced angiopoietin-2, an angiogenesis inhibitor. In contrast, prostate SMCs produced angiopoietin-1, a positive modulator of angiogenesis. Synthesis of the angiopoietins did not respond to androgen treatment. CONCLUSIONS: Prostate smooth muscle may play an important role in regulating vascular responses to androgen
— id: 35192, year: 2002, vol: 50, page: 83, stat: Journal Article,

Differentiation and stromal-induced growth promotion of murine prostatic tumors
Salm, Sarah N; Takao, Tetsuya; Tsujimura, Akira; Coetzee, Sandra; Moscatelli, David; Wilson, E Lynette
2002 May 15;51(3):175-188, Prostate
BACKGROUND: We have derived a panel of p53-null prostatic 'basal' and 'luminal' epithelial cell lines and their ras transformed counterparts to study stromal/epithelial interactions and the properties of tumors arising from 'basal' and 'luminal' cells. METHODS: Previously derived normal murine prostatic 'basal' epithelial (PE-B-1) and 'luminal' epithelial (PE-L-1) cell lines were transformed with N-Ras. These lines and a spontaneously transformed 'luminal' cell line were inoculated subcutaneously or orthotopically into athymic mice, alone or in combination with normal prostatic smooth muscle cells (SMC). RESULTS: All transformed lines formed subcutaneous tumors. SMC significantly enhanced the growth rate of the tumors arising from the 'basal' and one of the 'luminal' cell lines. The transformed 'basal' line gave rise to tumors expressing both 'basal' and 'luminal' cytokeratins. CONCLUSIONS: Prostatic SMC promote the growth of transformed epithelial cells, suggesting that prostatic stroma may promote tumor development. Furthermore, transformed 'basal' cells give rise to tumors containing 'luminal' cells, suggesting that although most human tumors have a 'luminal' phenotype, they may originate from transformed 'basal' cells
— id: 35191, year: 2002, vol: 51, page: 175, stat: Journal Article,

Proximal location of mouse prostate epithelial stem cells: a model of prostatic homeostasis
Tsujimura, Akira; Koikawa, Yasuhiro; Salm, Sarah; Takao, Tetsuya; Coetzee, Sandra; Moscatelli, David; Shapiro, Ellen; Lepor, Herbert; Sun, Tung-Tien; Wilson, E Lynette
2002 Jun 24;157(7):1257-1265, Journal of cell biology
Stem cells are believed to regulate normal prostatic homeostasis and to play a role in the etiology of prostate cancer and benign prostatic hyperplasia. We show here that the proximal region of mouse prostatic ducts is enriched in a subpopulation of epithelial cells that exhibit three important attributes of epithelial stem cells: they are slow cycling, possess a high in vitro proliferative potential, and can reconstitute highly branched glandular ductal structures in collagen gels. We propose a model of prostatic homeostasis in which mouse prostatic epithelial stem cells are concentrated in the proximal region of prostatic ducts while the transit-amplifying cells occupy the distal region of the ducts. This model can account for many biological differences between cells of the proximal and distal regions, and has implications for prostatic disease formation
— id: 32485, year: 2002, vol: 157, page: 1257, stat: Journal Article,

Generation of active TGF-beta by prostatic cell cocultures using novel basal and luminal prostatic epithelial cell lines
Salm SN; Koikawa Y; Ogilvie V; Tsujimura A; Coetzee S; Moscatelli D; Moore E; Lepor H; Shapiro E; Sun TT; Wilson EL
2000 Jul;184(1):70-79, Journal of cellular physiology
Two prostatic epithelial lines, one of basal origin and one of luminal origin, were established from the dorsolateral prostates of p53 null mice. The cell lines are nontumorigenic when inoculated subcutaneously under the renal capsule or intraprostatically in syngeneic mice. The luminal cell line (PE-L-1) expresses cytokeratins 8 and 18 and the basal cell line (PE-B-1) expresses cytokeratins 5 and 14. The basal cells require serum for growth, whereas the luminal cells grow only in serum-free medium. Both cell lines require the presence of growth factors for optimal growth in culture, with EGF and FGF-2 having the greatest effect on the growth rate. Both lines express androgen receptor (AR) mRNA and protein. Androgen stimulates growth of the basal cell line, indicating that the ARs are functional, whereas growth of the luminal cells is unaffected by androgens. The luminal line is significantly inhibited by exogenous TGF-beta and produces low levels of endogenous TGF-beta. In contrast, the basal cell line produces significant amounts of TGF-beta and its growth is not influenced by this cytokine. Coculture of luminal cells with prostatic smooth muscle cells results in the generation of increased levels of biologically active TGF-beta, indicating a paracrine mechanism of TGF-beta activation that may be involved in the maintenance of normal prostatic function. To our knowledge this is the first report describing both basal and luminal prostatic cell lines from a single inbred animal species and the first indication that prostatic epithelial and stromal cells interact to generate the biologically active form of TGF-beta. These lines will provide an important model for determining basal/luminal interactions in both in vitro and in vivo assays.
— id: 11685, year: 2000, vol: 184, page: 70, stat: Journal Article,

Transforming growth factor-beta is an autocrine mitogen for a novel androgen-responsive murine prostatic smooth muscle cell line, PSMC1
Salm SN; Koikawa Y; Ogilvie V; Tsujimura A; Coetzee S; Moscatelli D; Moore E; Lepor H; Shapiro E; Sun TT; Wilson EL
2000 Dec;185(3):416-424, Journal of cellular physiology
A prostatic smooth muscle cell line (PSMC1) was established from the dorsolateral prostate of p53 null mice. The cell line is nontumorigenic when inoculated subcutaneously, under the renal capsule or intraprostatically in syngeneic mice. These cells express alpha-smooth muscle actin (alpha-SMA), indicating their smooth muscle origin, and TGF-beta significantly enhances expression of alpha-SMA. The cells express both androgen receptor (AR) mRNA and protein, and respond mitogenically to physiological concentrations of androgens. PSMC1 cells produce significant amounts of TGF-beta, which stimulates growth by an autocrine mechanism. Dihydrotestosterone (DHT) increases proliferation of PSMC1 cells by promoting TGF-beta secretion. Considering the significant inhibitory effect of TGF-beta on prostatic epithelial cells and its stimulatory effect on the PSMC1 cells, we postulate that TGF-beta produced by prostatic smooth muscle cells may have a paracrine effect on the prostatic epithelium. We also postulate that TGF-beta may be involved in the etiology of benign prostatic hyperplasia (BPH) by stimulating excessive stromal proliferation. Line PSMC1 is the first reported androgen-responsive murine smooth muscle cell line. It will be useful for in vivo and in vitro experiments to study the mechanisms of androgen action on prostatic stroma and for delineating the interactions that occur between prostatic smooth muscle and epithelium that may lead to prostatic diseases such as BPH
— id: 26907, year: 2000, vol: 185, page: 416, stat: Journal Article,

Endothelial and primitive hematopoietic cell surface markers are co-expressed on a CD34+population that expresses fibroblast growth factor receptors (FGFRs)
Burger, PE; Coetzee, S; Salm, S; Cook, P; Fan, Y; McKeehan, WL; Kan, M; Suda, T; Hebbel, RP; Novitsky, N; Wilson, EL
1999 NOV 15 ;94(10):464A-464A, Blood
— id: 54779, year: 1999, vol: 94, page: 464A, stat: Journal Article,

Cellular glycosylphosphatidylinositol-specific phospholipase D regulates urokinase receptor shedding and cell surface expression
Wilhelm OG; Wilhelm S; Escott GM; Lutz V; Magdolen V; Schmitt M; Rifkin DB; Wilson EL; Graeff H; Brunner G
1999 Aug;180(2):225-235, Journal of cellular physiology
The glycosylphosphatidylinositol (GPI)-anchored, multifunctional receptor for the serine proteinase, urokinase plasminogen activator (uPAR, CD87), regulates plasminogen activation and cell migration, adhesion, and proliferation. uPAR occurs in functionally distinct, membrane-anchored and soluble isoforms (s-uPAR) in vitro and in vivo. Recent evidence indicates that s-uPAR present in the circulation of cancer patients correlates with tumor malignancy and represents a valuable prognostic marker in certain types of cancer. We have therefore analyzed the mechanism of uPAR shedding in vitro. We present evidence that uPAR is actively released from ovarian cancer cells since the rate of receptor shedding did not correlate with uPAR expression. While s-uPAR was derived from the cell surface, it lacked the hydrophobic portion of the GPI moiety indicating anchor cleavage. We show that uPAR release is catalyzed by cellular GPI-specific phospholipase D (GPI-PLD), an enzyme cleaving the GPI anchor of the receptor. Thus, recombinant GPI-PLD expression increased receptor release up to fourfold. Conversely, a 40% reduction in GPI-PLD activity by GPI-PLD antisense mRNA expression inhibited uPAR release by more than 60%. We found that GPI-PLD also regulated uPAR expression, possibly by releasing a GPI-anchored growth factor. Our data suggest that cellular GPI-PLD might be involved in the generation of circulating prognostic markers in cancer and possibly regulate the function of GPI-anchored proteins by generating functionally distinct, soluble counterparts
— id: 35194, year: 1999, vol: 180, page: 225, stat: Journal Article,

Isolation and characterization of a CD34+ population that expresses fibroblast growth factor receptors
Burger, PE; Coetzee, S; Cook, P; Fan, Y; McKeehan, WL; Kan, M; Suda, T; Mansveldt, E; Novitsky, N; Wilson, EL
1998 NOV 15 ;92(10):56A-56A, Blood
— id: 53626, year: 1998, vol: 92, page: 56A, stat: Journal Article,

bFGF and TGF-beta have antagonistic effects on c-kit expression in K562 cells
Burger, PE; Buhring, HJ; Gowans, A; Wilson, EL
1997 NOV 15 ;90(10):3520-3520, Blood
— id: 53134, year: 1997, vol: 90, page: 3520, stat: Journal Article,

Enhanced expression of fibroblast growth factor receptors (FGF-Rs) on K562 cells during the S/G2/M phase of the cell cycle compared to the G1 phase
Coetzee, S; Hirst, J; McKeehan, WL; Kan, M; Burger, PE; Wilson, EL
1997 NOV 15 ;90(10):1409-1409, Blood
— id: 53145, year: 1997, vol: 90, page: 1409, stat: Journal Article,

Urokinase-type plasminogen activator-deficient mice are predisposed to staphylococcal botryomycosis, pleuritis, and effacement of lymphoid follicles
Shapiro RL; Duquette JG; Nunes I; Roses DF; Harris MN; Wilson EL; Rifkin DB
1997 Jan;150(1):359-369, American journal of pathology
Urokinase-type plasminogen activator (uPA) is thought to be an important mediator in the proteolytic degradation of extracellular matrix components observed in a wide variety of normal physiological and pathological conditions. However, the phenotype of a recently developed strain of urokinase-deficient (uPA-/-) mice appears to be normal when maintained under ideal nonstressful conditions. We report an outbreak of botryomycosis, an unusual staphylococcal infection, in a colony of uPA-deficient mice. A detailed histological examination of these uPA-deficient animals also revealed a variety of previously unreported phenotypic abnormalities such as pleuritis and the effacement of lymphoid follicles in the regional lymph nodes and spleen. Additional phenotypic abnormalities such as dystrophic calcifications and rectal prolapse were also observed in the uPA-deficient population. These abnormalities were also noted in ostensibly healthy uPA-deficient animals. Botryomycosis did not affect a colony of wild-type (uPA+/+) animals maintained concurrently under identical conditions in the same room. The peculiar predisposition of the uPA-deficient animals to this rare bacterial infection and the development of phenotypic abnormalities associated with the targeted disruption the uPA gene suggests that uPA contributes significantly to the cutaneous microenvironment and is additional evidence of the extensive involvement of the plasminogen activators in mammalian physiology
— id: 12426, year: 1997, vol: 150, page: 359, stat: Journal Article,

Inhibition of glycosylphosphatidylinositol (GPI) phospholipase D by suramin-like compounds
Brunner G; Zalkow L; Burgess E; Rifkin DB; Wilson EL; Gruszecka-Kowalik E; Powis G
1996 Sep-Oct;16(5A):2513-2516, Anticancer research
A number of proteins are found attached to the plasma membrane of mammalian cells by a glycosylphosphatidylinositol (GPI) anchor that can be cleaved by GPI specific phospholipase D (GPI-PLD). There are no known specific inhibitors of GPI-PLD. We examined some inhibitors of phosphatidylinositol specific phospholipase C (PI-PLC) for their ability to inhibit human serum and human bone marrow cell GPI-PLD. Azo analogues of suramin were found to be potent inhibitors of GPI-PLD. One compound had an IC50 of 3.7 microM that was 10-fold lower than the IC50 required to inhibit PI-PLC. The azo suramin analogues inhibited cancer cell growth at concentrations similar to those required to inhibit GPI-PLD, and below concentrations required to inhibit growth factor binding. It is possible that inhibition of cell growth might be related to the ability of the compounds to inhibit GPI-PLD
— id: 12546, year: 1996, vol: 16, page: 2513, stat: Journal Article,

Fibroblast growth factors promote a primitive phenotype in K562 cells
Burger, PE; Wilson, EL
1996 NOV 15 ;88(10):3179-3179, Blood
— id: 52696, year: 1996, vol: 88, page: 3179, stat: Journal Article,

The regulation of TGF-beta activity by hematopoietic cells
Coetzee, S; Burger, P; Rifkin, D; Wilson, EL
1996 NOV 15 ;88(10):2150-2150, Blood
— id: 52712, year: 1996, vol: 88, page: 2150, stat: Journal Article,

Elevated intracellular level of basic fibroblast growth factor correlates with stage of chronic lymphocytic leukemia and is associated with resistance to fludarabine
Menzel T; Rahman Z; Calleja E; White K; Wilson EL; Wieder R; Gabrilove J
1996 Feb 1;87(3):1056-1063, Blood
Chronic lymphocytic leukemia (CLL) is characterized by delayed senescence and slow accumulation of monoclonal, small lymphocytes. Basic fibroblast growth factor (bFGF) is a pleiotropic cytokine that plays a role in hematopoiesis and apoptosis. Elevated bFGF levels have been detected in urine from patients with a variety of neoplastic diseases including various leukemias; however, the cellular source of the bFGF has not been determined. In this study, the intracellular bFGF level in lymphocytes of 36 patients with B-CLL and 15 normal donors was determined using an enzyme-linked immunoassay. In cells derived from patients with high-risk disease, the median level of intracellular bFGF was 381.5 pg/2 x 10(5) cells, compared with a median of 90.5 pg/2 x 10(5) cells in patients with intermediate disease. In patients with low-risk disease, the median bFGF level was 4.9 pg/2 x 10(5) cells, and in normal controls, it was 6.0 pg/2 x 10(5) cells. The difference in the bFGF levels was significant for the comparison between low- and intermediate-risk (P = .00119), low- and high-risk (P < .0001), and intermediate- and high-risk disease (P = .0001). Immunofluorescent stains of peripheral blood mononuclear cells confirmed CLL lymphocytes as a cellular source of bFGF. To evaluate the potential contribution of elevated intracellular bFGF levels to the phenotype of CLL cells, leukemic cells were cultured in vitro with an apoptotic stimulus (fludarabine). CLL cells with high intracellular levels of bFGF appeared to be more resistant to fludarabine treatment. The addition of bFGF to fludarabine-treated CLL cells resulted in a delay of apoptosis and prolonged survival. These data suggest that bFGF may contribute to the resistance of CLL cells to an apoptotic stimulus
— id: 35195, year: 1996, vol: 87, page: 1056, stat: Journal Article,

Induction of primary cutaneous melanocytic neoplasms in urokinase-type plasminogen activator (uPA)-deficient and wild-type mice: cellular blue nevi invade but do not progress to malignant melanoma in uPA-deficient animals
Shapiro RL; Duquette JG; Roses DF; Nunes I; Harris MN; Kamino H; Wilson EL; Rifkin DB
1996 Aug 1;56(15):3597-3604, Cancer research
Evidence suggests that the plasminogen activators (PAs), in particular urokinase-type PA (uPA), play a pivotal role in tumor invasion and metastasis. We studied the contribution of the PAs to the malignant phenotype through the chemical induction of melanocytic neoplasms in uPA-deficient mice. Primary tumors were induced and promoted concurrently in 35 uPA-/- deficient and 35 uPA+/+ wild-type mice using a single application of 7,12-dimethylbenz(a)anthracene followed by repetitive applications of croton oil. Animals were sacrificed at 60-day intervals for 1 year. At necropsy, the four largest pigmented lesions in each animal were excised, characterized histologically, and evaluated microscopically for evidence of invasion. The regional lymph nodes, lungs, and solid abdominal visceral organs were sectioned and examined microscopically for evidence of metastatic disease. Cellular blue nevi were induced in 100% of uPA-/- and uPA+/+ promoted animals. Although a reduction in the radial and vertical progression of these lesions was noted in the uPA-deficient mice compared with the wild-type group, more than 95% of cellular blue nevi induced in both groups of animals invaded the underlying tissues. These lesions did not metastasize to the regional lymph nodes. Malignant melanoma arose in 5 of 35 (14.3%) of promoted wild-type mice. These tumors were locally aggressive, produced tissue-type PA, but were not metastatic to the regional nodes, lungs, or abdominal viscera. These results indicate that the invasive capability of melanocytic lesions may depend more on tissue-type PA than uPA activity. No melanomas were induced in the uPA-/- mice. The resistance of the uPA -/- strain to melanoma induction suggests that uPA contributes to malignant progression. We propose that the absence of uPA negatively affects tumorigenesis by decreasing the liberation and availability of growth factors such as basic fibroblast growth factor
— id: 12575, year: 1996, vol: 56, page: 3597, stat: Journal Article,

bFGF and TGF-beta exert antagonistic effects on erythroid differentiation in K562 cells
Burger, PE; Lukey, PT; Wilson, EL
1995 NOV 15 ;86(10):560-560, Blood
— id: 129599, year: 1995, vol: 86, page: 560, stat: Journal Article,

Basic fibroblast growth factor (bFGF) transfected stromal cells promote the growth of Dami leukemic cells
Ogilvie, V; Coetzee, S; Rifkin, D; Quesenberry, P; Wilson, EL
1995 NOV 15 ;86(10):1229-1229, Blood
— id: 53118, year: 1995, vol: 86, page: 1229, stat: Journal Article,

An endogenous glycosylphosphatidylinositol-specific phospholipase D releases basic fibroblast growth factor-heparan sulfate proteoglycan complexes from human bone marrow cultures
Brunner G; Metz CN; Nguyen H; Gabrilove J; Patel SR; Davitz MA; Rifkin DB; Wilson EL
1994 Apr 15;83(8):2115-2125, Blood
Basic fibroblast growth factor (bFGF) is a hematopoietic cytokine that stimulates stromal and stem cell growth. It binds to a glycosylphosphatidylinositol (GPI)-anchored heparan sulfate proteoglycan on human bone marrow (BM) stromal cells. The bFGF-proteoglycan complex is biologically active and is released by addition of exogenous phosphatidylinositol-specific phospholipase C. In this study, we show the presence of an endogenous GPI-specific phospholipase D (GPI-PLD) that releases the bFGF-binding heparan sulfate proteoglycan and the variant surface glycoprotein (a model GPI-anchored protein) from BM cultures. An involvement of proteases in this process is unlikely, because released proteoglycan contained the GPI anchor component, ethanol-amine, and protease inhibitors did not diminish the release. The mechanism of release is likely to involve a GPI-PLD and not a GPI-specific phospholipase C, because the release of variant surface glycoprotein did not reveal an epitope called the cross-reacting determinant that is exposed by phospholipase C-catalyzed GPI anchor cleavage. In addition, phosphatidic acid (which is specifically a product of GPI-PLD-catalyzed anchor cleavage) was generated during the spontaneous release of the GPI-anchored variant surface glycoprotein. We also detected GPI-PLD-specific enzyme activity and mRNA in BM cells. Therefore, we conclude that an endogenous GPI-PLD releases bFGF-heparan sulfate proteoglycan complexes from human BM cultures. This mechanism of GPI anchor cleavage could be relevant for mobilizing biologically active bFGF in BM. An endogenous GPI-PLD could also release other GPI-anchored proteins important for hematopoiesis and other physiologic processes
— id: 56502, year: 1994, vol: 83, page: 2115, stat: Journal Article,

GROWTH-FACTOR REGULATION OF THE GLYCOSYLPHOSPHATIDYLINOSITOL (GPI)-SPECIFIC PHOSPHOLIPASE-D IN BONE-MARROW STROMAL CELLS
BRUNNER, G; GULATI, M; GABRILOVE, JL; RIFKIN, DB; WILSON, EL
1994 NOV 15 ;84(10):A564-A564, Blood
— id: 52291, year: 1994, vol: 84, page: A564, stat: Journal Article,

Basic fibroblast growth factor antagonizes transforming growth factor beta-mediated erythroid differentiation in K562 cells
Burger PE; Dowdle EB; Lukey PT; Wilson EL
1994 Apr 1;83(7):1808-1812, Blood
Basic fibroblast growth factor (bFGF) and transforming growth factor-beta 1 (TGF-beta) have both been shown to act on hematopoietic progenitor cells. bFGF is a hematopoietic cytokine that acts on progenitor cells in concert with other cytokines to promote their proliferation. TGF-beta induces erythroid differentiation in K562 cells. To determine whether bFGF might act on progenitor cells by antagonizing the effects of cytokines that induce differentiation, we determined the effects of bFGF on the TGF-beta-mediated induction of hemoglobin synthesis in K562 cells. bFGF antagonized the TGF-beta-mediated induction of hemoglobin in a dose-dependent manner, with 0.1 ng/mL bFGF inhibiting hemoglobin induction by 40% and 10 ng/mL bFGF completely abrogating hemoglobin production. bFGF was most effective at antagonizing the TGF-beta-mediated induction of hemoglobin if it and TGF-beta were added simultaneously to K562 cells, but delayed addition of bFGF to TGF-beta-treated cultures still resulted in significant inhibition of hemoglobin synthesis. The inhibitory effects of bFGF on hemoglobin production were fully reversible, showing that bFGF did not permanently alter the phenotype of K562 cells. The hemin-mediated induction of hemoglobin synthesis in K562 cells was only partially negated by bFGF. bFGF also diminished the expression of glycophorin A on the surface of K562 cells. These results indicate that bFGF might increase progenitor/stem cell numbers by antagonizing the effects of cytokines that induce differentiation, thereby increasing the pool of proliferating progenitor/stem cells
— id: 57512, year: 1994, vol: 83, page: 1808, stat: Journal Article,

BASIC FIBROBLAST GROWTH-FACTOR DECREASES EXPRESSION OF GLYCOPHORIN-A ON K562 CELLS
BURGER, PE; LUKEY, PT; WILSON EL
1994 NOV 15 ;84(10):A420-A420, Blood
— id: 129598, year: 1994, vol: 84, page: A420, stat: Journal Article,

BASIC FIBROBLAST GROWTH-FACTOR (BFGF) TRANSFECTED STROMAL CELLS SUPPORT SURVIVAL OF MO7E CELLS
COETZEE, S; OGILVIE, V; BRUNNER, G; QUARTO, N; RIFKIN, D; QUESENBERRY, P; WILSON, EL
1994 NOV 15 ;84(10):A279-A279, Blood
— id: 52284, year: 1994, vol: 84, page: A279, stat: Journal Article,

Stem cell factor and basic fibroblast growth factor are synergistic in augmenting committed myeloid progenitor cell growth
Gabrilove JL; White K; Rahman Z; Wilson EL
1994 Feb 15;83(4):907-910, Blood
Stem cell factor (SCF) and basic fibroblast growth factor (bFGF) are hematopoietic cytokines produced by bone marrow stromal cells. It is known that, although SCF and bFGF have limited clonogenic activity on their own, they can augment colony-stimulating factor (CSF)-mediated progenitor cell growth. Because these factors are both sequestered by stromal cells, we examined their interaction on progenitor cell growth in conjunction with granulocyte-macrophage-CSF (GM-CSF). In this study, we show that clonogenic growth derived from low-density bone marrow cells stimulated by GM-CSF is significantly augmented (P < .001) in the presence of maximal (100 ng/mL) concentrations of SCF in combination with 100 ng/mL of bFGF. When CD34+ cells are used, the synergistic effect of bFGF and SCF for GM-CSF-mediated progenitor cell growth is further increased, resulting in as much as a sevenfold increase in detectable colony-forming units granulocyte-macrophage (P < .001). These data suggest that the synergistic activity of bFGF and SCF is mediated directly on hematopoietic precursors. These observations suggest that bFGF and SCF, concentrated locally on stromal cell surfaces, might interact in concert with other hematopoietic cytokines to regulate stem cell proliferation and differentiation in hematopoietic niches in the bone marrow
— id: 57424, year: 1994, vol: 83, page: 907, stat: Journal Article,

Release of GPI-anchored membrane proteins by a cell-associated GPI-specific phospholipase D
Metz CN; Brunner G; Choi-Miura NH; Nguyen H; Gabrilove J; Caras IW; Altszuler N; Rifkin DB; Wilson EL; Davitz MA
1994 Apr 1;13(7):1741-1751, EMBO journal
Although many glycosylphosphatidylinositol (GPI)-anchored proteins have been observed as soluble forms, the mechanisms by which they are released from the cell surface have not been demonstrated. We show here that a cell-associated GPI-specific phospholipase D (GPI-PLD) releases the GPI-anchored, complement regulatory protein decay-accelerating factor (DAF) from HeLa cells, as well as the basic fibroblast growth factor-binding heparan sulfate proteoglycan from bone marrow stromal cells. DAF found in the HeLa cell culture supernatants contained both [3H]ethanolamine and [3H]inositol, but not [3H]palmitic acid, whereas the soluble heparan sulfate proteoglycan present in bone marrow stromal cell culture supernatants contained [3H]ethanolamine. 125I-labeled GPI-DAF incorporated into the plasma membranes of these two cell types was released in a soluble form lacking the fatty acid GPI-anchor component. GPI-PLD activity was detected in lysates of both HeLa and bone marrow stromal cells. Treatment of HeLa cells with 1,10-phenanthroline, an inhibitor of GPI-PLD, reduced the release of [3H]ethanolamine-DAF by 70%. The hydrolysis of these GPI-anchored molecules is likely to be mediated by an endogenous GPI-PLD because [3H]ethanolamine DAF is constitutively released from HeLa cells maintained in serum-free medium. Furthermore, using PCR, a GPI-PLD mRNA has been identified in cDNA libraries prepared from both cell types. These studies are the first demonstration of the physiologically relevant release of GPI-anchored proteins from cells by a GPI-PLD
— id: 7885, year: 1994, vol: 13, page: 1741, stat: Journal Article,

Mechanism of action of angiostatic steroids: suppression of plasminogen activator activity via stimulation of plasminogen activator inhibitor synthesis
Blei F; Wilson EL; Mignatti P; Rifkin DB
1993 Jun;155(3):568-578, Journal of cellular physiology
Recently, a novel class of angiostatic steroids which block angiogenesis in several systems has been described. Since the elaboration of proteases is believed to be an important component of angiogenesis, we tested whether these steroids blocked the fibrinolytic response of endothelial cells to the angiogenic protein, basic fibroblast growth factor [bFGF]). Cultured bovine aortic endothelial (BAE) cells were incubated with bFGF and/or medroxyprogesterone acetate (MPA), an angiostatic steroid which has been shown to inhibit vascularization, collagenolysis, and tumor growth. When bFGF (3 ng/ml) was added to confluent monolayers of BAE cells, plasminogen activator (PA) activity in the medium was increased threefold. In contrast, MPA at 10(-6) M, 10(-7) M, 10(-8) M, and 10(-9) M decreased PA levels in the medium by 83%, 83%, 75%, and 39%, respectively. The stimulation of PA levels in BAE cells by bFGF (3 ng/ml) was abrogated by the presence of 10(-6) M MPA. This decrease in PA activity was found to be mediated by a significant increase in plasminogen activator inhibitor type-1 (PAI-1) production. MPA, therefore, negated one of the important enzymatic activities associated with the angiogenic process. In contrast to the decreased levels of secreted PA in cultures exposed simultaneously to MPA and bFGF, cell-associated PA levels remained high, consistent with earlier observations indicating that PAI-1 does not inhibit cell-associated PA. Thus, angiostatic steroids may exert their inhibitory effects on angiogenesis by increasing the synthesis of PAI-1. This, in turn, inhibits PA activity and, therefore, plasmin generation, which is essential for the invasive aspect of angiogenesis
— id: 8234, year: 1993, vol: 155, page: 568, stat: Journal Article,

Basic fibroblast growth factor expression in human bone marrow and peripheral blood cells
Brunner G; Nguyen H; Gabrilove J; Rifkin DB; Wilson EL
1993 Feb 1;81(3):631-638, Blood
We have shown previously that basic fibroblast growth factor (bFGF) is a mitogen for human bone marrow (BM) stromal cells and that bFGF stimulates myelopoiesis in primary BM cultures. In this article, we demonstrate the presence of bFGF in two cell lineages in human BM and peripheral blood as well as the deposition of bFGF into the extracellular matrix of BM stromal cell cultures. In immunofluorescence experiments on BM and peripheral blood smears, megakaryocytes and platelets stained strongly for bFGF, whereas weaker staining was observed in immature and mature cells of the granulocyte series. The presence of bFGF in platelets was confirmed by enzyme-linked immunosorbent assay as well as by immunoprecipitation followed by immunoblotting. bFGF was synthesized by BM stromal cell cultures and was found either cell associated or localized in the nucleus and the nucleoli, and its location was dependent on the fixation procedure used. Addition of exogenous bFGF to stromal cells showed the presence of extracellular binding molecules for this cytokine. bFGF could be released from these sites by soluble heparin or phosphatidylinositol-specific phospholipase C. This study supports the role of bFGF as a stromal cell mitogen and stimulator of myelopoiesis. The data indicate that the stromal cells produce bFGF and that their extracellular matrix can serve as a reservoir for this growth factor. In addition, the results suggest a possible involvement of bFGF in platelet function as well as in megakaryocytopoiesis
— id: 13270, year: 1993, vol: 81, page: 631, stat: Journal Article,

MOBILIZATION OF BFGF-PROTEOGLYCAN COMPLEXES IN HUMAN BONE-MARROW CULTURES BY A GPI-SPECIFIC PHOSPHOLIPASE-D
BRUNNER, G; NGUYEN, H; RIFKIN, DB; GABRILOVE, J; WILSON, EL
1993 NOV 15 ;82(10):A370-A370, Blood
— id: 52146, year: 1993, vol: 82, page: A370, stat: Journal Article,

Basic fibroblast growth factor promotes the proliferation of human megakaryocyte progenitor cells
Bruno E; Cooper RJ; Wilson EL; Gabrilove JL; Hoffman R
1993 Jul 15;82(2):430-435, Blood
Basic fibroblast growth factor (bFGF), a multifunctional growth factor produced by bone marrow stromal cells, is known to be a potent modulator of hematopoiesis. Because bFGF is present in both human megakaryocytes (MKs) and platelets, we have hypothesized that this growth factor might affect human megakaryocytopoiesis. To test this hypothesis, either low density bone marrow (BM) cells (LDBM), a human BM subpopulation (CD34+ DR+) enriched for the colony-forming unit megakaryocyte (CFU-MK) or a BM subpopulation (CD34+ DR-) enriched for the more primitive burst-forming unit megakaryocyte (BFU-MK) were assayed in the presence of this growth factor. The effect of bFGF on MK colony formation differed according to the cell population assayed. bFGF alone had on MK colony-stimulating activity (MK-CSA) when either CD34+ DR+ or CD34+ DR- BM cells were cloned, but exhibited MK-CSA equivalent to that of interleukin-3 (IL-3) when LDBM cells were used as the target cell population. The MK-CSA of bFGF was inhibited by the addition of neutralizing antisera to either IL-3 and/or granulocyte-macrophage colony-stimulating factor (GM-CSF) but not IL-6. The addition of excess amounts of either IL-3 or GM-CSF to cultures containing bFGF plus anti-IL-3 or anti-GM-CSF reversed the inhibition by the corresponding antisera. The addition of bFGF and IL-3 to assays containing CD34+ DR+ or CD34+ DR- cells increased the size of both CFU-MK- and BFU-MK-derived colonies, respectively, when compared with assays containing IL-3 alone. This increase in MK colony size mediated by bFGF was not affected by addition of either an anti-GM-CSF or anti-IL-6 neutralizing antisera. When LDBM cells were assayed, bFGF alone increased CFU-MK-derived colony size when compared with control values. However, this potentiation of MK colony size by bFGF could be reversed by the addition of either anti-IL-3 or anti-GM-CSF but not anti-IL-6 antisera. In addition, the effects of bFGF and IL-3 on the size of MK colonies cloned from LDBM were not additive. These results suggest that bFGF affects human megakaryocytopoiesis by directly promoting MK progenitor cell proliferation and stimulating BM accessory cells to release growth factor(s) with MK-CSA, such as IL-3 and GM-CSF. We conclude that bFGF, likely produced by cellular components of the BM microenvironment, plays an important role in the control of human megakaryocytopoiesis
— id: 35196, year: 1993, vol: 82, page: 430, stat: Journal Article,

ROLE OF HEPARAN-SULFATE IN MODULATING HUMAN HEMATOPOIETIC STEM-CELL ATTACHMENT TO CYTOKINES AND EXTRACELLULAR-MATRIX MOLECULES
BRUNO, E; GUSCAR, TK; LUIKART, SD; WILSON, EL; DIXIT, V; LONG, MW; HOFFMAN, R
1993 NOV 15 ;82(10):A21-A21, Blood
— id: 52139, year: 1993, vol: 82, page: A21, stat: Journal Article,

BASIC FIBROBLAST GROWTH-FACTOR ANTAGONIZES TRANSFORMING GROWTH-FACTOR BETA-MEDIATED ERYTHROID-DIFFERENTIATION IN K562 CELLS
BURGER, PE; DOWDLE, EB; WILSON, EL
1993 NOV 15 ;82(10):A99-A99, Blood
— id: 52140, year: 1993, vol: 82, page: A99, stat: Journal Article,

BASIC FIBROBLAST GROWTH-FACTOR STIMULATES PROLIFERATION OF MURINE BONE-MARROW STROMAL CELLS IN THE ABSENCE OF SERUM
COETZEE, S; BRUNNER, G; MOSCATELLI, D; QUESENBERRY, P; WILSON, EL
1993 NOV 15 ;82(10):A496-A496, Blood
— id: 52148, year: 1993, vol: 82, page: A496, stat: Journal Article,

BASIC FIBROBLAST GROWTH-FACTOR AUGMENTS THE GM-CSF-DEPENDENT GROWTH OF MO7 CELLS
COETZEE, S; MOSCATELLI, D; LIUZZO, J; GABRILOVE, J; WILSON, EL
1993 NOV 15 ;82(10):A236-A236, Blood
— id: 52144, year: 1993, vol: 82, page: A236, stat: Journal Article,

Basic fibroblast growth factor counteracts the suppressive effect of transforming growth factor-beta 1 on human myeloid progenitor cells
Gabrilove JL; Wong G; Bollenbacher E; White K; Kojima S; Wilson EL
1993 Feb 15;81(4):909-915, Blood
We have previously shown that basic fibroblast growth factor (bFGF) is mitogenic for human bone marrow stromal cells and enhances myelopoiesis in human long-term bone marrow culture. In the present study, we examined the mechanism by which bFGF enhances granulopoiesis. We observed that bFGF significantly abrogated the inhibitory effect of transforming growth factor-beta 1 (TGF-beta 1) on granulocyte-macrophage colony-stimulating factor (GM-CSF)-supported progenitor cell growth (P = .009). The partial reversal of TGF-beta 1-mediated suppression was dependent on the dose of bFGF used. In addition, we noted that the inclusion of neutralizing antibody to TGF-beta 1 significantly augmented the clonogenic response to GM-CSF. We have also shown that 10 ng/mL or 100 ng/mL of bFGF resulted in a 30% to 100% increase in GM-CSF-mediated progenitor cell growth (P = .0001). These data suggest that bFGF may enhance myelopoiesis by modulating the inhibitory response to TGF-beta 1
— id: 35197, year: 1993, vol: 81, page: 909, stat: Journal Article,

PHOSPHOLIPASE-C RELEASE OF BIOLOGICALLY-ACTIVE BASIC FIBROBLAST GROWTH FACTOR-HEPARAN SULFATE PROTEOGLYCAN COMPLEXES FROM HUMAN BONE-MARROW CULTURES
BRUNNER, G; GABRILOVE, J; RIFKIN, DB; WILSON, EL
1992 JAN ;20(1):107-107, Experimental hematology
— id: 52124, year: 1992, vol: 20, page: 107, stat: Journal Article,

RECOMBINANT HUMAN BASIC FIBROBLAST GROWTH-FACTOR AFFECTS THE PROLIFERATION OF HUMAN MEGAKARYOCYTE PROGENITOR CELLS
BRUNO, E; COOPER, RJ; WILSON, EL; GABRILOVE, JL; HOFFMAN, R
1992 JUL ;20(6):823-823, Experimental hematology
— id: 51935, year: 1992, vol: 20, page: 823, stat: Journal Article,

Unusual growth characteristics of human melanoma xenografts in the nude mouse: a model for desmoplasia, dormancy and progression
Gartner MF; Fearns C; Wilson EL; Campbell JA; Dowdle EB
1992 Apr;65(4):487-490, British journal of cancer
When human melanoma cells are injected into nude mice they usually give rise to tumours that grow progressively and do not elicit a prominent host response. We have recently developed a melanoma cell line, UCT-Mel 7, that did not show these characteristics. In the first place UCT-Mel 7 showed a consistently unusual, phasic growth pattern. After a short initial period of limited growth (phase 1), the tumour ceased growing and remained static for 2-3 months (phase 2). The tumour then regressed (phase 3) to enter a second period of quiescence (phase 4) which was eventually broken by the emergence of a rapidly growing lethal tumour (phase 5). Of particular interest was the fact that the rate at which the tumours grew correlated closely with their collagen content. During the prolonged, phase 2 plateau, the tumours were intensely desmoplastic; rapidly growing phase 5 tumours, that had escaped from dormancy, contained very little collagen and virtually no reticulin. This cell line helps to fill an important need for an experimental system for the study of desmoplasia, dormancy and progression
— id: 35199, year: 1992, vol: 65, page: 487, stat: Journal Article,

Fibroblast-dependent tumorigenicity of melanoma xenografts in athymic mice
Gartner MF; Wilson EL; Dowdle EB
1992 Jul 9;51(5):788-791, International journal of cancer
Two human melanoma cell lines, UCT-Mel 2 and UCT-Mel 3, were invariably tumorigenic in nude mice when inoculated s.c. in doses of 10(6) cells or higher; 10(5) cells or less did not give rise to tumours. In this report we show that otherwise sub-tumorigenic inocula developed into vigorously growing tumour xenografts when co-inoculated with normal fibroblasts. Fibroblasts derived from adult, neonatal or embryonic tissues all functioned as complementing cells, as did cells of human or murine origin. There was, however, a requirement for complementing cell viability, since ethanol-killed fibroblasts were inefficacious. The fibroblast effect was dose-dependent and was not observed if injections of fibroblasts and melanoma cells were separated anatomically or temporally. We have shown, by titrating admixtures of melanoma cells and fibroblasts, that fibroblasts are, in quantitative terms, more efficacious than melanoma cells as complementing cells. The system we describe provides a useful model for the study of stromal-cell regulation of tumour growth
— id: 35198, year: 1992, vol: 51, page: 788, stat: Journal Article,

Regulation of proteolytic activity in human bone marrow stromal cells by basic fibroblast growth factor, interleukin-1, and transforming growth factor beta
Hannocks MJ; Oliver L; Gabrilove JL; Wilson EL
1992 Mar 1;79(5):1178-1184, Blood
Plasminogen activators (PAs) and/or plasmin may be involved in hematopoietic regulation. These enzymes release biologically relevant cytokines such as basic fibroblast growth factor (bFGF) from matrix and cell surfaces. In addition, transforming growth factor beta (TGF beta) and interleukin-1 beta (IL-1 beta) are converted from inactive to active forms by plasmin. Therefore, we studied the regulation of PAs and their specific inhibitors, PA inhibitor 1 (PAI-1) and PA inhibitor 2 (PAI-2), in human bone marrow stromal fibroblasts by IL-1 beta, bFGF, and TGF beta. All three cytokines stimulated PA secretion. IL-1 beta at 10(4) U/mL increased urokinase (u-PA) levels approximately 10-fold, bFGF at 0.2 ng/mL also increased production 10-fold, but increased predominantly tissue PA (t-PA) expression. TGF beta at 0.2 ng/mL increased u-PA production up to 300-fold. PAI-1 and PAI-2 are also regulated by these cytokines. IL-1 beta decreased PAI-1 levels by 50% and stimulated PAI-2 levels sixfold. bFGF had minimal effects on PAI-1 and TGF beta increased PAI-1 levels twofold. Neither of these agents had an effect on PAI-2 levels. Thus, three cytokines relevant to bone marrow physiology regulate PA and inhibitor production by human bone marrow stromal fibroblasts. In this manner PA and plasmin generation in specific microenvironments in the bone marrow may be one of the factors orchestrating the complex series of events, which results in an efficient exquisitely regulated hematopoietic process
— id: 13681, year: 1992, vol: 79, page: 1178, stat: Journal Article,

THE REGULATION OF PLASMINOGEN-ACTIVATOR ACTIVITY IN HUMAN BONE-MARROW STROMAL CELLS
HANNOCKS, MJ; RIFKIN, DB; OLIVER, L; GABRILOVE, J; WILSON, EL
1992 JAN ;20(1):108-108, Experimental hematology
— id: 52125, year: 1992, vol: 20, page: 108, stat: Journal Article,

SECRETION OF PLASMINOGEN ACTIVATORS BY NORMAL BONE-MARROW CELLS AND LEUKEMIC MYELOID CELLS
WILSON, EL; JACOBS, P; FRANCIS, GE; OLIVER, L; BURGER, P; DOWDLE, EB
1992 JAN ;6(3):77-79, Fibrinolysis
The secretion of tissue plasminogen activator (t-PA) and urokinase by normal human bone marrow cells is a differentiation linked property with t-PA being produced by primitive progenitor cells and urokinase being produced by more differentiated cells and by mature neutrophils and macrophages. Cells from patients with acute myeloid leukaemia also secrete both types of plasminogen activator (PA) and the type of enzyme secreted has prognostic significance. Patients whose cells secrete t-PA die rapidly and fail chemotherapy whereas 80% of those individuals whose cells secrete urokinase enter remission following chemotherapy. The generation of plasmin in the haemopoietic microenvironment would influence haemopoiesis by converting precursor cytokines to active species and would also release various haemopoietic cytokines from cell surfaces and matrix facilitating their interaction with cell surface receptors. The inappropriate secretion of PAs by leukaemic cells could result in abnormal haemopoiesis due to the aberrant plasmin-mediated activation and release of various cytokine species
— id: 52104, year: 1992, vol: 6, page: 77, stat: Journal Article,

ANGIOSTATIC STEROIDS INHIBIT ENDOTHELIAL-CELL PLASMINOGEN-ACTIVATOR ACTIVITY
BLEI, F; WILSON, EL; RIFKIN, DB
1991 APR ;29(4):A137-A137, Pediatric research
— id: 51662, year: 1991, vol: 29, page: A137, stat: Journal Article,

Phospholipase C release of basic fibroblast growth factor from human bone marrow cultures as a biologically active complex with a phosphatidylinositol-anchored heparan sulfate proteoglycan
Brunner G; Gabrilove J; Rifkin DB; Wilson EL
1991 Sep;114(6):1275-1283, Journal of cell biology
Basic fibroblast growth factor (bFGF) is a potent mitogen for human bone marrow stromal cells and stimulates haematopoiesis in vitro. We report here that primary human bone marrow cultures contain bFGF and express heparin-like bFGF binding sites on the cell surface and in the extracellular matrix (ECM). bFGF bound predominantly to a 200-kD cell surface heparan sulfate proteoglycan (HSPG), which was also found in conditioned medium. bFGF was released from bone marrow cultures by incubation with phosphatidylinositol-specific phospholipase C (PI-PLC) and, less efficiently, by plasmin. Solubilized bFGF was found as a complex with the 200-kD HSPG. The complex was biologically active as shown by its ability to stimulate plasminogen activator production in bovine aortic endothelial cells. bFGF-HSPG complexes of bovine endothelial cells, however, were not released by PI-PLC. While only trace amounts of the bFGF-binding 200-kD HSPG were released spontaneously from bone marrow cultures, incubation with PI-PLC solubilized almost all of the 200-kD HSPG. The HSPG could be metabolically labeled with ethanolamine or palmitate, which was partially removed by treatment with PI-PLC. These findings indicate linkage of the HSPG to the cell surface via a phosphatidylinositol anchor. Plasmin released the 200-kD HSPG less efficiently than PI-PLC. We conclude that HSPGs of human bone marrow serve as a reservoir for bFGF, from which it can be released in a biologically active form via a dual mechanism; one involving a putative endogenous phospholipase, the other involving the proteolytic cascade of plasminogen activation
— id: 13933, year: 1991, vol: 114, page: 1275, stat: Journal Article,

Inhibition of urokinase-type plasminogen activator by antibodies: the effect on dissemination of a human tumor in the nude mouse
Ossowski L; Russo-Payne H; Wilson EL
1991 Jan 1;51(1):274-281, Cancer research
Nude mice given inoculations s.c. of a human squamous carcinoma--HEp3 (1.5 x 10(6) cells/mouse)--developed invasive tumors that produced high levels of urokinase-type plasminogen activator (uPA) and metastasized predictably to the lungs and lymph nodes of the host. To investigate the role of uPA in invasion and metastasis, mice given inoculations of tumor cells were treated daily with s.c. injections of specific, anti-human uPA antibodies (rabbit polyclonal, 150 inhibitory units; mouse monoclonal, 3000 inhibitory units/mouse/day). Control mice received either saline or preimmune rabbit immunoglobulins. A total of approximately 50 mice was studied. The tumors were surgically excised 10 to 17 days postinoculation when weighing 1 to 2 g. Antibody administration was discontinued after tumor excision. Two strategies were used: (a) following the removal of tumors the mice were maintained and observed until respiratory distress, indicative of lung metastasis, was evident; or (b) their lungs were examined for evidence of metastasis on the day of tumor removal. While histological sections of s.c. tumors excised from control mice indicated extensive local invasion, evidence of invasion was absent in most tumors excised from mice in which tumor uPA was inhibited by the antibody (P less than 0.025). The inhibition of local invasion did not, however, lead to a reduced incidence of distant metastasis. Since we found that the presence of HEp3 tumors in mice elicits a pronounced granulocytosis, we propose that this response may facilitate the spread of tumor cells by a mechanism independent of endogenous tumor proteases
— id: 35200, year: 1991, vol: 51, page: 274, stat: Journal Article,

Basic fibroblast growth factor stimulates myelopoiesis in long-term human bone marrow cultures
Wilson EL; Rifkin DB; Kelly F; Hannocks MJ; Gabrilove JL
1991 Mar 1;77(5):954-960, Blood
We previously showed that basic fibroblast growth factor (bFGF) is a potent mitogen for human bone marrow (BM) stromal cells and significantly delays their senescence. In the present study, we demonstrated that low concentrations of bFGF (0.2 to 2 ng/mL) enhance myelopoiesis in long-term human BM culture. Addition of bFGF to long-term BM cultures resulted in an increase in (a) the number of nonadherent cells (sixfold), particularly those of the neutrophil granulocyte series; (b) the number of nonadherent granulocyte colony-stimulating factor (G-CSF)- and granulocyte-macrophage colony-stimulating factor (GM-CSF)-responsive progenitor cells; (c) the number of adherent foci of hematopoietic cells (10-fold); and (d) the number of progenitor cells in the adherent stromal cell layer. These effects were not noted with higher concentrations of bFGF (20 ng/mL). Thus, low concentrations of bFGF effectively augment myelopoiesis in human long-term BM cultures, and bFGF may therefore be a regulator of the hematopoietic system in vitro and in vivo
— id: 57423, year: 1991, vol: 77, page: 954, stat: Journal Article,

Chemotherapy of adult acute nonlymphoblastic leukaemia
Jacobs P; Martell RW; Wilson EL
1990 ;23(1):27-40, Haematologia (Budapest)
Seventy-two consecutive and previously untreated adults with acute non-lymphoblastic leukaemia (ANLL), having a median age of 36 years (range 12 to 71), were prospectively randomised to receive conventional doses of cytosine arabinoside and doxorubicin combined with either etoposide (CTR III) or 6-thioguanine (DAT). Morbidity was comparable between the two regimens and complete remission (CR) rates of 52% and 62% respectively (p greater than 0.50) were not influenced by age above or below 50 years, initial white cell count, French-American-British classification, or race. However, growth pattern in the GM: CFUc assay was found to identify a subgroup of patients who had a significantly higher CR rate. Similarly, the secretion of tissue plasminogen activator by leukaemic blasts in vitro uniformly predicted for primary drug resistance, whereas a CR rate of 68% was associated with production of the urokinase type or a mixture of both enzymes. Remission duration and survival did not differ between these two forms of chemotherapy, nor were they influenced by immunotherapy with C. parvum or the duration of maintenance therapy, whereas age below 50 and the species of plasminogen activator secreted were significant prognostic factors. It is concluded that etoposide can be substituted for 6-thioguanine in these cytosine arabinoside and doxorubicin-containing regimens and that for both combinations the most sensitive prognostic factor for CR and survival is the species of plasminogen activator secreted in vitro by the leukaemic blasts
— id: 35204, year: 1990, vol: 23, page: 27, stat: Journal Article,

A prospective analysis of prognostic factors in the myelodysplastic syndromes
Knottenbelt E; Jacobs P; Wilson EL
1990 Jan 20;77(2):69-74, South African medical journal
In view of the uncertainty regarding the natural history and management of individuals with the myelodysplastic syndromes, a prospective study of 43 consecutive and previously untreated patients was undertaken in order to identify haematological features that could predict for poor prognosis. A significant correlation between percentage of blasts in the bone marrow, maturity index and the number of cell lineages involved was demonstrated with both the risk of leukaemic transformation and survival. It remains to be determined whether further accumulation of data will result in similar predictive values for karyotypic analysis, in vitro bone marrow culture and the species of plasminogen activator secreted by the cells. Since treatment ranges from red cell transfusion and administration of maturation-inducing agents to aggressive cytotoxic chemotherapy or bone marrow transplantation, the development of predictive models, based on relevant prognostic factors, remains the most rational basis for choices between these various options
— id: 35202, year: 1990, vol: 77, page: 69, stat: Journal Article,

Long-term culture of human bone marrow stromal cells in the presence of basic fibroblast growth factor
Oliver LJ; Rifkin DB; Gabrilove J; Hannocks MJ; Wilson EL
1990 ;3(3):231-236, Growth factors
Basic fibroblast growth factor (bFGF) is a potent mitogen for human bone marrow stromal cells. Normally, large numbers of human bone marrow stromal cells are difficult to obtain. However, nanogram/ml concentrations of bFGF stimulate the growth of passaged bone marrow stromal cells both in media formulated for optimal growth of stromal cells and in a simple mixture of RPMI-1640 and 10% fetal calf serum facilitating the successive expansion of stromal cells through multiple passages. bFGF also greatly accelerates the formation of a primary stromal cell layer following inoculation of newly harvested bone marrow cells into dishes. In the presence of bFGF, the stromal cells attain high densities, lose their contact inhibition and grow in multilayered sheets. Heparin greatly potentiates the stimulatory effect of low concentrations of bFGF. The effects of bFGF are fully reversible: cells cultured in the presence of this factor for multiple passages revert to normal growth rates following trypsinization and subculture. A short (4 h) exposure of the cells to bFGF elicits profound growth stimulation. This supports the hypothesis that this factor binds to glycosaminoglycans in the cell matrix which act as a storage reservoir for this cytokine
— id: 35203, year: 1990, vol: 3, page: 231, stat: Journal Article,

Regulation and secretion of plasminogen activators and their inhibitors in a human leukemic cell line (K562)
Oliver LJ; Keeton M; Wilson EL
1989 Sep;74(4):1321-1327, Blood
The secretion of tissue plasminogen activator (t-PA), urokinase (u-PA) and their inhibitors by the human leukemia cell line K562 was examined. K562 cells normally secrete both t-PA and u-PA in a ratio of 3:1. After addition of 10 or 1 ng/mL phorbol myristate acetate (PMA) to K562 cells, a marked decrease in enzymatic activity is observed in the medium. However, when t-PA antigen rather than activity is measured, an increased amount is found in the medium under these conditions. PMA also induces secretion of the two inhibitors of plasminogen activator: plasminogen activator inhibitor 1 (PAI-1) and plasminogen activator inhibitor 2 (PAI-2). This accounts for the decrease in total enzymatic activity under conditions when production of t-PA antigen is increased. A study of the time course of induction revealed that the synthesis of plasminogen activator occurred before that of its inhibitors. Low concentrations of PMA (0.1 ng/mL) induce t-PA antigen primarily and not the inhibitors. This results in an increase in total enzymatic activity, with 94% of the secreted activity being t-PA. Thus, the secretion of plasminogen activators and their inhibitors can be manipulated in certain leukemic cells by inducers such as PMA
— id: 10518, year: 1989, vol: 74, page: 1321, stat: Journal Article,

Abortion
Wilson EL
1989 Summer;7(2):55-58, Health matrix
PIP: If you are pregnant and near 40 years old there is 1/137 chance that your child may have Down's syndrome, or 1/65 chance he will have a physical or mental problem. There are tests that can indicate these problems but they increase the risk of spontaneous abortion. A woman should not be forced to carry an unwanted child, and the needs of childless couples should not be addressed in abortion discussions. The Roe v. Wade case made the distinction of not having to determine when life begins, but when it can be sustained outside the body. The Missouri statute states that human life begins at conception, an unborn child has protectable life interests and the parents of that child have protectable life interests of the unborn child in relation to life, health and its well being. States that are really concerned with the interests of unborn children should improve prenatal care, educate teens on contraception, AIDS, and be concerned about violent behavior and smoking. Voters in Michigan and Arkansas approved a law to stop the use of public funds for abortion, other than saving the mother's life. Pro- choice advocates are concerned that the conservative appointees to the supreme court will reverse the previous decision. O
— id: 35201, year: 1989, vol: 7, page: 55, stat: Journal Article,

Metastasis of a human melanoma cell line in the nude mouse
Wilson EL; Gartner MF; Campbell JA; Dowdle EB
1988 Jan 15;41(1):83-86, International journal of cancer
A human melanoma cell line has been established which when inoculated subcutaneously into nude mice, is consistently metastatic. In order to document blood-borne spread, it was necessary to excise the primary tumour so prolonging the life of the animal and allowing metastases to become apparent. Macroscopically detectable metastatic spread at autopsy was reliably indicated by weight loss of the animals. Metastases were widespread and involved the lungs, abdominal cavity and organs and the gonads. The size of the primary tumour at the time of its removal, and not the period of s.c. growth, determined the incidence of metastatic disease. Removal of tumours weighing less than 0.6 g prevented metastasis, whereas all of the animals showed widely disseminated disease if the tumour was allowed to attain a size of 1.6 g before excision
— id: 35205, year: 1988, vol: 41, page: 83, stat: Journal Article,

Evidence for the involvement of DNA topoisomerase II in neutrophil-granulocyte differentiation
Francis GE; Berney JJ; North PS; Khan Z; Wilson EL; Jacobs P; Ali M
1987 Sep;1(9):653-659, Leukemia
Agents that slow cellular proliferation usually stimulate myeloid differentiation. The demonstration in this report of an anomalous inhibitory behavior of the epipodophyllotoxin VP16-213, an agent known to inhibit the enzyme DNA topoisomerase II, prompted us to investigate the role of this enzyme in both changes in DNA supercoiling and in DNA strand breakage and reunion events occurring during the induction of neutrophil-granulocyte differentiation. We recently reported that retinoic acid, an inducer of granulocytic differentiation, stimulates transient relaxation of DNA supercoiling. We now show that this is associated with the formation of small numbers of protein-linked DNA breaks (a characteristic of topoisomerase reactions). Both events are perturbed by VP16-213, and since this agent inhibits subsequent differentiation, these observations raise the possibility of a role for DNA topoisomerase II in granulocytic differentiation. The possible relevance of these findings to mechanisms of leukemogenesis is discussed
— id: 35207, year: 1987, vol: 1, page: 653, stat: Journal Article,

Growth of a human carcinoma (HEp3) in nude mice: rapid and efficient metastasis
Ossowski L; Russo H; Gartner M; Wilson EL
1987 Nov;133(2):288-296, Journal of cellular physiology
Our aim was to identify conditions which would permit the development of spontaneous metastasis of a human tumor in nude mice in a rapid and predictable manner and to explore ways to quantitate metastasis. Using a human squamous carcinoma--HEp3--we determined that invasiveness and metastasis were influenced by the host. HEp3 cells grew very rapidly and without a significant lag period in Balb/c and NIH(S)-II nude mice kept in aseptic conditions; a much longer lag period was observed in NIH-Swiss mice kept in conventional conditions. The HEp3 tumor displayed a highly invasive behavior in N-NIH(S)-II mice, in which it invaded the body wall, gaining access to the peritoneal cavity. Microinvasion was noted in all strains of mice inoculated with HEp3 cells. To prolong survival of the mice until metastases became evident, primary tumors were excised when they weighed 1-2 gm. N-NIH(S)-II and Balb/c nude mice, maintained in germ-free conditions, were most receptive to the development of metastases-lung metastases developed in 80% of these mice. Over 60% of all metastases were present within 4 weeks following the removal of the primary. Only 26% of tumor bearing NIH-Swiss developed lung metastases. Lung metastases were observed in some mice in the absence of local microinvasion, local tumor recurrence, and regardless of the presence of lymph node involvement. They were also noted in mice from which primary tumors were not excised. We compared three methods of lung metastasis detection: histology, detection of tumor cells in the cultures of lung minces, and the measurement of the levels of human urokinase-type plasminogen activator directly in the lysates of lungs. The detection of tumor cells in cultures of lung minces appeared to be the most sensitive of these methods and the determination of enzyme in lung lysates seemed to hold most promise for a quantitative approach. These data indicate that, the type of tumor, as well as the genetic background and the maintenance conditions of the host, have to be carefully selected to ensure the successful outcome of the particular tumor-host interaction being studied. Adherence to these guidelines allowed us, in the case of the HEp3 tumor, to develop a rapid, predictable, and efficient model in which to study factors affecting metastasis of this human tumor
— id: 35206, year: 1987, vol: 133, page: 288, stat: Journal Article,

Differentiation-linked secretion of urokinase and tissue plasminogen activator by normal human hemopoietic cells
Wilson EL; Francis GE
1987 Jun 1;165(6):1609-1623, Journal of experimental medicine
Previous studies have shown that the response of patients with acute myeloid leukemia to induction chemotherapy can be predicted by the species of plasminogen activator that their cells secrete. Patients whose cells secreted tissue plasminogen activator (tPA) only failed to respond to combination chemotherapy. Individuals whose leukemic cells display features of the early progenitor phenotype also respond poorly to therapy. This suggested that the two species of plasminogen activator secreted by leukemic cells might be produced by normal cells at distinct stages of differentiation. These results indicate that the secretion of the two enzyme types is a differentiation-linked property of normal cells with tPA being produced by granulocyte/macrophage progenitors and urokinase by more differentiated cells and by mature neutrophils and macrophages
— id: 35208, year: 1987, vol: 165, page: 1609, stat: Journal Article,

The secretion of plasminogen activators by human bone marrow progenitor cells
Wilson EL; Francis GE
1987 ;31(6):181-184, Haematology & blood transfusion
— id: 35209, year: 1987, vol: 31, page: 181, stat: Journal Article,

Hormonal regulation of extracellular plasminogen activators and Mr approximately 54,000 plasminogen activator inhibitor in human neoplastic cell lines, studied with monoclonal antibodies
Andreasen PA; Christensen TH; Huang JY; Nielsen LS; Wilson EL; Dano K
1986 May;45(2-3):137-147, Molecular & cellular endocrinology
We have studied the regulation by glucocorticoids and dibutyryl cAMP of the amounts of urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA) and a Mr approximately 54000 plasminogen activator inhibitor accumulated in serum-free conditioned culture fluid by a human fibrosarcoma, a human glioblastoma and a human melanoma cell line (HT-1080, UCT/gl-1 and Bowes). For the quantitation of u-PA and t-PA, we used sandwich-type ELISA with a combination of polyclonal and monoclonal antibodies. For an estimation of variations in the amount of the inhibitor, we used sodium dodecyl sulphate-polyacrylamide gel electrophoresis followed by Coomassie blue staining of conditioned culture fluid proteins, the inhibitor protein band being identified by its selective removal by passage of the conditioned culture fluids through a column with monoclonal antibodies against the inhibitor. The modulation of the 3 proteins by the hormonal agents varied greatly between the cell lines. The proteins were independently regulated, in the sense that the hormonal agents did not concomitantly change their levels in the direction expected either to increase or decrease total extracellular plasminogen activator activity. In conditioned culture fluids containing both t-PA and inhibitor, the two were present in the medium as a Mr approximately 120 000 complex. In contrast, no u-PA inhibitor complexes were found in conditioned culture fluid from any of the cell lines; this is likely to be due to the occurrence of u-PA in the culture fluid in the one-chain proenzyme form, which, unlike active u-PA, does not react with the inhibitor. These findings illustrate the complexity of the regulation of extracellular plasminogen activator activity, and imply that the presumed functional diversity of u-PA and t-PA may be related to their independent regulation
— id: 35211, year: 1986, vol: 45, page: 137, stat: Journal Article,

Characterization of seven human melanoma cell lines: melanogenesis and secretion of plasminogen activators
Hoal-Van Helden EG; Wilson EL; Dowdle EB
1986 Aug;54(2):287-295, British journal of cancer
Permanent cell lines (UCT-Mel 1 through 7) were established from biopsies of metastatic tissue taken from seven patients with malignant melanoma. Cells from these lines were all aneuploid and all grew as non-contact-inhibited, adherent monolayers. All of the lines, with the remarkable exception of UCT-Mel 6, formed tumours in nude mice, expressed the melanoma M-18 antigen and synthesized plasminogen activators exclusively of the tissue-type. UCT-Mel 6 cells were non tumourigenic, they lacked the M-18 antigen and they synthesized plasminogen activators exclusively of the urokinase type. UCT-Mel 1 and UCT-Mel 2 formed pigment in vitro and both of these lines showed an increase in pigment content and tyrosinase synthesis with increasing cell density. The rate of plasminogen activator released by UCT-Mel 1 and UCT-Mel 3 declined strikingly as the cells became confluent. Assuming that proteolytic activity is required for cell migration in vivo; that tyrosinase synthesis reflects expression of the differentiated phenotype and that melanoma cells retain some of the characteristics of neural crest cells, we suggest that the effects of confluence and close cell-cell contact provide a useful experimental counterpart for the study of normal neural crest all behaviour that is characterized by an inverse relationship between migration and a protease secretion on the one hand and pigmentation on the other
— id: 35210, year: 1986, vol: 54, page: 287, stat: Journal Article,

Plasminogen activator as a prognostic factor in hematological malignancies
Wilson EL; Jacobs P; Oliver L
1985 ;29(2-3):197-199, Haematology & blood transfusion
— id: 35212, year: 1985, vol: 29, page: 197, stat: Journal Article,

The regulation of tissue plasminogen activator activity by human fibroblasts
Hoal EG; Wilson EL; Dowdle EB
1983 Aug;34(1):273-279, Cell
We have found that live and ethanol-fixed fibroblasts, when covered with conditioned medium containing tissue plasminogen activator, associate with the enzyme and remove it from the medium. Binding of tissue plasminogen activator to fixed cells showed equilibrium kinetics with maximal uptake corresponding to 2.4 units of enzyme per 10(6) fixed cells. Enzyme bound to fixed cells could activate plasminogen and produce plaques of caseinolysis in casein-plasminogen-agar overlays. Electrophoretic analysis showed it covalently attached to a fibroblast component with a molecular weight of 40,000-50,000. Sequestration of tissue plasminogen activator by live fibroblasts showed nonsaturable first order kinetics with a rate constant of 0.465/hr. We conclude that active enzyme is bound to a surface receptor, then internalized and degraded. Fibroblasts did not release the binding molecule into the medium; binding of tissue plasminogen activator from the medium was unaffected by heparin or thrombin. This phenomenon differs from that described by Baker et al. and ascribed to 'proteasenexin.'
— id: 35213, year: 1983, vol: 34, page: 273, stat: Journal Article,

Secretion of plasminogen activators by human myeloid leukemic cells: modulation and therapeutic correlations
Wilson EL; Jacobs P; Dowdle EB
1983 ;28(3):78-80, Haematology & blood transfusion
— id: 35216, year: 1983, vol: 28, page: 78, stat: Journal Article,

The effects of dexamethasone and tetradecanoyl phorbol acetate on plasminogen activator release by human acute myeloid leukemia cells
Wilson EL; Jacobs P; Dowdle EB
1983 Mar;61(3):561-567, Blood
This investigation was undertaken to examine the extent to which leukemic cell functions are susceptible to regulation in vitro and to investigate their heterogeneity in this regard. Since plasminogen activator release is known to be a modulatable cellular function that can be influenced by antiinflammatory steroids and tetradecanoyl phorbol acetate (TPA), the effect of these two compounds on the secretion of urokinase- or tissue-type enzymes by leukemic cells was studied. The release of both enzyme species could be stimulated or suppressed by these substances by mechanisms that were inhibitable by actinomycin-D and hence required transcription of new mRNA. Plasminogen activator release by cells from 41/45 patients with AML was either stimulated or inhibited by 10(-7) M dexamethasone, implying that most AML cells possess glucocorticoid receptors. In 26/45 cases, the enzyme was inhibited by this steroid to less than 25% of control values. Pronounced inhibition of this degree was not encountered with normal polymorphonuclear leukocytes. Plasminogen activator secretion by AML cells was profoundly inhibited in 20/41 cases by 1 ng/ml TPA and stimulated in 8/41 cases. Leukemic blasts varied considerably in their response to dexamethasone and TPA. Plasminogen activator release should prove a sensitive means of monitoring the responses of AML cells to biologically active compounds
— id: 35215, year: 1983, vol: 61, page: 561, stat: Journal Article,

The secretion of plasminogen activators by human myeloid leukemic cells in vitro
Wilson EL; Jacobs P; Dowdle EB
1983 Mar;61(3):568-574, Blood
Peripheral blood cell preparation from 23 normal subjects and 72 patients with acute and 32 patients with chronic myeloid leukemia were cultured in vitro and released plasminogen activators were analyzed. The quantity of plasminogen activator secreted by leukemic cells varied widely and could not be correlated with the clinical severity of the disease. Immunochemical and electrophoretic techniques have been used to show that normal peripheral blood granulocytes released exclusively urokinase-like plasminogen activator, whereas leukemic cells secreted either urokinase or a tissue activator-like enzyme. The molecular species of enzyme released by acute myeloid leukemic cells may serve as a diagnostic marker of relevance to the management of this disease, since patients with acute myeloid leukemia whose cells released only tissue plasminogen activator did not respond to combination chemotherapy. Tissue plasminogen activators released by leukemic cells may display an unusual electrophoretic pattern that resembles that shown by urokinase. Immunochemical procedures are therefore essential for the correct identification of these enzymes
— id: 35214, year: 1983, vol: 61, page: 568, stat: Journal Article,

Plasminogen activating enzyme in cultured glioblastoma cells. An immunofluorescence study with monoclonal antibody
Dano K; Dabelsteen E; Nielsen LS; Kaltoft K; Wilson EL; Zeuthen J
1982 Nov;30(11):1165-1170, Journal of histochemistry & cytochemistry
A monoclonal antibody against a 52,000 dalton human plasminogen activating enzyme (HPA52) was used for immunofluorescence staining of cultured glioblastoma cells. The fluorescence was located in the cytoplasm of the cells. A pronounced variation in the staining intensity was observed between the individual cells. The specificity of the fluorescent stain was supported by the findings that 1) no staining was obtained with a monoclonal antibody of the same subclass, but with irrelevant specificity (anti-2,4,6-trinitrophenyl); 2) adsorption with HPA52 purified to homogeneity removed the ability of anti-HPA52 to mediate staining; 3) the glioblastoma cells contained HPA52, as measured by enzymatic assay, while melanoma cells that were not stained did not contain HPA52 activity; 4) dexamethasone reduced both the enzymatically determined HPA52 content and the immunofluorescence in parallel, while progesterone affected none of these parameters; 5) we have previously found that culture fluid conditioned by the glioblastoma cells apart from HPA52 does not contain detectable amounts of any protein that binds to anti-HPA52. Several advantages of immunohistochemical detection of plasminogen activators compared with enzyme histochemical methods are discussed, among these that the immunohistochemical method distinguishes between plasminogen activators of different types
— id: 35219, year: 1982, vol: 30, page: 1165, stat: Journal Article,

Variable effects of retinoids on two pigmenting human melanoma cell lines
Hoal E; Wilson EL; Dowdle EB
1982 Dec;42(12):5191-5195, Cancer research
Two melanoma cell lines, each derived from a different patient with metastatic disease, were very similar in their appearance, their growth characteristics, and their tendency to differentiate and to pigment in culture as they become confluent. These lines, UCT-Mel 1 and UCT-Mel 2, were used to study the effects of retinoic acid and other derivatives of vitamin A. When added to UCT-Mel 1 cells, retinoids had only a modest effect on plasminogen activator release and were without measurable effect on morphology, growth, or tyrosinase synthesis. In contrast, when added to UCT-Mel 2 cells, retinoids appeared to induce a more differentiated state evident as an inhibition of cell proliferation and the assumption of a dendritic morphology. Paradoxically, however, retinoids caused a striking inhibition of the density-dependent intracellular accumulation of tyrosinase and melanin that was taken to represent spontaneous in vitro differentiation. Culture of UCT-Mel 2 cells in the presence of retinoic acid resulted in initial inhibition followed by marked stimulation of cellular plasminogen activator release. The data suggest that the manner in which retinoids exert their effects on cells in vitro does not depend on the histological origin of the tumor cells being studied but on the innate responsiveness of that particular cell line to the retinoid or compound in question
— id: 35218, year: 1982, vol: 42, page: 5191, stat: Journal Article,

Purification of zymogen to plasminogen activator from human glioblastoma cells by affinity chromatography with monoclonal antibody
Nielsen LS; Hansen JG; Skriver L; Wilson EL; Kaltoft K; Zeuthen J; Dano K
1982 Dec 7;21(25):6410-6415, Biochemistry
Incorporation of the serine protease active site reagent diisopropyl fluorophosphate (DFP) into a plasminogen activator with an Mr of approximately 52000 released from cultured human glioblastoma cells was strongly enhanced by incubation with plasmin. This observation led to the isolation of an inactive form of the enzyme from serum-free conditioned culture fluid by affinity chromatography on a column of a Sepharose-bound monoclonal antibody raised against urokinase. An 831-fold purification was obtained with a yield of 41%. The purified molecule was homogeneous as evaluated by polyacrylamide gel electrophoresis with sodium dodecyl sulfate (NaDodSO4), having one stainable band under nonreducing as well as reducing conditions with an Mr of approximately 52000. It was unable to activate plasminogen, but catalytic amounts of plasmin converted it into active enzyme. After NaDodSO4-polyacrylamide gel electrophoresis, the active enzyme showed one band under nonreducing conditions, but after reduction, two bands with Mr values of approximately 20000 and 32000 were observed. The active enzyme incorporated [3H]DFP into the approximately Mr 32000 band, while no incorporation was observed into the inactive form. These findings show that the Mr 52000 human plasminogen activator exists in a proenzyme form consisting of a single polypeptide chain that by proteolysis between half-cystine residues is converted into the active enzyme consisting of two chains with molecular weights of approximately 20000 and 32000, the active site being on the latter chain. The results are consistent with the active form of the enzyme being identical with the higher molecular weight form of urokinase, and together with recent observations that a murine plasminogen activator is released from sarcoma virus transformed cells as an inactive proenzyme, they suggest that zymogens to plasminogen activators are of more general occurrence
— id: 35217, year: 1982, vol: 21, page: 6410, stat: Journal Article,

Colorimetric assay of urinary 5-hydroxy-3-indoleacetic acid
Shihabi ZK; Wilson EL
1982 Apr;15(2):106-108, Clinical biochemistry
We describe a method for 5-hydroxy-3-indoleacetic acid in urine based on its reaction with nitrosonaphthol in the presence of mercaptoethanolamine, an odorless compound. Mercaptoethanolamine shifts the color maximum from 540 nm to 640 nm, intensifies the color and discharges the color of interfering substances. The method is rapid and free from interferences
— id: 35220, year: 1982, vol: 15, page: 106, stat: Journal Article,

Challenges in preventive medicine and public health: the health status of refugees from Indochina
Flynn JP; Sorley DL; Wilson EL
1980 May;29(5):60-63, Maryland state medical journal
— id: 35222, year: 1980, vol: 29, page: 60, stat: Journal Article,

Molecular species of plasminogen activators secreted by normal and neoplastic human cells
Wilson EL; Becker ML; Hoal EG; Dowdle EB
1980 Mar;40(3):933-938, Cancer research
A survey of 56 normal and neoplastic tissues has shown that plasminogen activators were released by cultured human cells in several molecular weights and their susceptibility to inhibition by antibodies to the normal urinary enzyme, urokinase. Melanoma cells characteristically secreted plasminogen activators which were immunochemically distinct from urokinase and which migrated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a prominent, closely spaced doublet with an apparent molecular weight of approximately 70,000 and a minor molecular weight component of approximately 60,000. Enzymes with similar characteristics have been observed in serum-free harvest fluids collected from other neoplastic tissue (a breast carcinoma, a glioblastoma, a malignant teratoma, a uterine sarcoma, and a carcinoma of the renal pelvis) and from normal tissue (8-week embryo fibroblasts, normal esophageal fibroblasts, and one culture of normal adult bladder epithelium). Plasminogen activators released by cells derived from most normal adult tissues, or from a 26-week-old embryo, and from other tumors of ectodermal or mesenchymal origin were inhibited by anti-urokinase antibody and showed a closely spaced doublet with a molecular weight of 60,000 as the most abundant molecular species with no evidence of the enzyme with a molecular weight of 70,000
— id: 35223, year: 1980, vol: 40, page: 933, stat: Journal Article,

Effects of retinoids on normal and neoplastic human cells cultured in vitro
Wilson EL; Dowdle EB
1980 Dec;40(12):4817-4820, Cancer research
The effects of retinoic acid on cultured human cells derived from normal and neoplastic tissues were studied. Retinoic acid consistently induced plasminogen activator synthesis by cells of mesenchymal origin (with the exception of adult skin fibroblasts) but not by cells of epithelial origin. The effect of retinoic acid was more pronounced than that of equimolar concentrations of retinol or retinyl acetate. Dexamethasone inhibited the retinoid-induced increase in plasminogen activator in lung- and foreskin-derived fibroblasts. Cells derived from normal or neoplastic tissues showed no consistent differences either in baseline rates of plasminogen activator release or in the magnitude of the retinoid effect
— id: 35221, year: 1980, vol: 40, page: 4817, stat: Journal Article,

Effect of vitamin A on plasminogen activator synthesis by chick embryo fibroblasts
Wilson EL; Reich E
1979 ;23(5):319-323, Haematology & blood transfusion
Low concentrations of Vitamin A stimulated plasminogen activator synthesis (PA) in chick embryo fibroblasts (CEF). It caused a dose dependent and reversible increase in PA synthesis in both normal CEF and CEF infected with a temperature sensitive mutant of Rous Sarcoma virus (RSV-Ts68). Both induction and deinduction of PA could be inhibited by Actinomycin D. Vitamin A also accentuated the morphological changes associated with transformation in the Rous Sarcoma virus infected cells. The effects of Vitamin A on PA synthesis were essentially similar to those of the known tumour promoter, phorbol myristate acetate (PMA). Both Vitamin A and PMA were found to act synergistically with sarcoma gene expression as far as PA synthesis was concerned
— id: 35225, year: 1979, vol: 23, page: 319, stat: Journal Article,

Modulation of plasminogen activator synthesis in chick embryo fibroblasts by cyclic nucleotides and phorobol myristate acetate
Wilson EL; Reich E
1979 May;39(5):1579-1586, Cancer research
To explore the interaction of tumor promoters and sarcoma virus transformation with cellular regulatory mechanisms, we have studied induction of plasminogen activator synthesis by these agents in a background of changing cyclic nucleotide concentrations. We have confirmed the original report of Wigler and Weinstein (Nature, 259: 232, 1976) that phorbol-12-myristate-13-acetate (PMA), a potent tumor promoter, induces high levels of plasminogen activator production by chick embryo fibroblasts. Sarcoma virus transformation sensitizes the fibroblasts by lowering the threshold concentration for response to the action of PMA, and the effects of transformation and PMA on plasminogen activator synthesis are synergistic rather than additive. The plasminogen activators produced in the PMA-, virus-induced, or synergistically stimulated cultures are indistinguishable. Enzyme production in all three conditions is strongly but reversibly inhibited when cyclic nucleotide levels are raised by exposure to cyclic adenosine-3':5'-monophosphate or cholera toxin. A substantial fraction of the morphological effect that accompanies transformation is not affected by concentrations of cyclic nucleotides that suppress plasminogen activator production, and the two phenomena are therefore at least partially independent expressions of transformation in this system
— id: 35224, year: 1979, vol: 39, page: 1579, stat: Journal Article,

Secretion of plasminogen activator by normal, reactive and neoplastic human tissues cultured in vitro
Wilson EL; Dowdle E
1978 Oct 15;22(4):390-399, International journal of cancer
Plasminogen activator secretion by 39 primary or early-passage cultures of malignant human neoplasms has been compared with that of 16 similar cultures of benign neoplasms and 39 cultures of normal or reactive tissue. While normal cells of mesenchymal or neural origin secreted considerably less plasminogen activator than did cells from frankly malignant tissues, elevated levels of enzyme secretion were also encountered in cultures of benign neoplastic or reactive cells. In the case of epithelial tissue, no consistent relationship between plasminogen activator secretion and neoplasia could be documented. Our failure to observe, for any particular cell type, a reproducible correlation between malignancy and plasminogen activator secretion may be attributable to the artificial conditions of in vitro culture, where normal in vivo regulatory mechanisms do not obtain
— id: 35226, year: 1978, vol: 22, page: 390, stat: Journal Article,

Plasminogen activator in chick fibroblasts: induction of synthesis by retinoic acid; synergism with viral transformation and phorbol ester
Wilson EL; Reich E
1978 Oct;15(2):385-392, Cell
This paper reports the effect of vitamin A and its derivatives, the retinoids, on plasminogen activator (PA) synthesis in chick embryo fibroblast cultures (CEF). Low concentrations of retinoic acid (RA) (10(-6)-10(-10) M) and the retinoids stimulated PA synthesis in CEF; the maximal stimulation achieved, 9--10 fold, was somewhat lower than that obtained with optimal concentrations of the potent tumor promoter phorbol myristate acetate (PMA). This action of RA required protein and mRNA synthesis but, in contrast to enzyme induction by PMA and/or sarcoma virus transformation, retinoid effects were not significantly inhibited by elevated concentrations of cAMP. In inducing and/or stimulating PA production, the effects of RA and sarcoma virus transformation were synergistic rather than additive. Analogous synergism was observed between RA and PMA, but only at suboptimal concentrations of the latter. RA did not affect PA production in normal or transformed cultures maximally stimulated by PMA. These findings may help to elucidate the role of retinoids in promoting tumor growth, tissue remodeling and teratogenesis
— id: 35227, year: 1978, vol: 15, page: 385, stat: Journal Article,

Evaluation of the use of resonant frequencies to characterize physical properties of human long bones
Dhoerty WP; Bovill EG; Wilson EL
1974 Nov;7(6):559-561, Journal of biomechanics
— id: 35228, year: 1974, vol: 7, page: 559, stat: Journal Article,

Beef-liver 5-aminolevulinic acid dehydratase. Purification and properties
Wilson EL; Burger PE; Dowdle EB
1972 Sep 25;29(3):563-571, European journal of biochemistry
— id: 35229, year: 1972, vol: 29, page: 563, stat: Journal Article,

Isolation and purification of ala-dehydratase
Wilson, E L; Dowdle, E B; Burger, P
1971 Sep 25;29(3):178-180, South African medical journal
— id: 35230, year: 1971, vol: 29, page: 178, stat: Journal Article,

Vaginal mucification induced by pituitary prolactin and placental lactogen in mice. A luteotrophic test for protein hormone preparations
Josimovich JB; Wilson EL; Leff A
1970 ;1(4):210-221, Gynecologic investigation
— id: 56386, year: 1970, vol: 1, page: 210, stat: Journal Article,