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  1. Article ; Online: Lymphangiogenesis Guidance Mechanisms and Therapeutic Implications in Pathological States of the Cornea.

    Patnam, Mehul / Dommaraju, Sunil R / Masood, Faisal / Herbst, Paula / Chang, Jin-Hong / Hu, Wen-Yang / Rosenblatt, Mark I / Azar, Dimitri T

    Cells

    2023  Volume 12, Issue 2

    Abstract: Corneal lymphangiogenesis is one component of the neovascularization observed in several inflammatory pathologies of the cornea including dry eye disease and corneal graft rejection. Following injury, corneal (lymph)angiogenic privilege is impaired, ... ...

    Abstract Corneal lymphangiogenesis is one component of the neovascularization observed in several inflammatory pathologies of the cornea including dry eye disease and corneal graft rejection. Following injury, corneal (lymph)angiogenic privilege is impaired, allowing ingrowth of blood and lymphatic vessels into the previously avascular cornea. While the mechanisms underlying pathological corneal hemangiogenesis have been well described, knowledge of the lymphangiogenesis guidance mechanisms in the cornea is relatively scarce. Various signaling pathways are involved in lymphangiogenesis guidance in general, each influencing one or multiple stages of lymphatic vessel development. Most endogenous factors that guide corneal lymphatic vessel growth or regression act via the vascular endothelial growth factor C signaling pathway, a central regulator of lymphangiogenesis. Several exogenous factors have recently been repurposed and shown to regulate corneal lymphangiogenesis, uncovering unique signaling pathways not previously known to influence lymphatic vessel guidance. A strong understanding of the relevant lymphangiogenesis guidance mechanisms can facilitate the development of targeted anti-lymphangiogenic therapeutics for corneal pathologies. In this review, we examine the current knowledge of lymphatic guidance cues, their regulation of inflammatory states in the cornea, and recently discovered anti-lymphangiogenic therapeutic modalities.
    MeSH term(s) Humans ; Lymphangiogenesis ; Corneal Neovascularization/drug therapy ; Corneal Neovascularization/metabolism ; Corneal Neovascularization/pathology ; Vascular Endothelial Growth Factor C/metabolism ; Cornea/metabolism ; Lymphatic Vessels/metabolism
    Chemical Substances Vascular Endothelial Growth Factor C
    Language English
    Publishing date 2023-01-14
    Publishing country Switzerland
    Document type Journal Article ; Review ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2661518-6
    ISSN 2073-4409 ; 2073-4409
    ISSN (online) 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells12020319
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Morphometric Analysis of Rat Prostate Development: Roles of MEK/ERK and Rho Signaling Pathways in Prostatic Morphogenesis.

    Hu, Wen-Yang / Afradiasbagharani, Parivash / Lu, Ranli / Liu, Lifeng / Birch, Lynn A / Prins, Gail S

    Biomolecules

    2021  Volume 11, Issue 12

    Abstract: The molecular mechanisms underlying prostate development can provide clues for prostate cancer research. It has been demonstrated that MEK/ERK signaling downstream of androgen-targeted FGF10 signaling directly induces prostatic branching during ... ...

    Abstract The molecular mechanisms underlying prostate development can provide clues for prostate cancer research. It has been demonstrated that MEK/ERK signaling downstream of androgen-targeted FGF10 signaling directly induces prostatic branching during development, while Rho/Rho-kinase can regulate prostate cell proliferation. MEK/ERK and Rho/Rho kinase regulate myosin light chain kinase (MLCK), and MLCK regulates myosin light chain phosphorylation (MLC-P), which is critical for cell fate, including cell proliferation, differentiation, and apoptosis. However, the roles and crosstalk of the MEK/ERK and Rho/Rho kinase signaling pathways in prostatic morphogenesis have not been examined. In the present study, we used numerical and image analysis to characterize lobe-specific rat prostatic branching during postnatal organ culture and investigated the roles of FGF10-MEK/ERK and Rho/Rho kinase signaling pathways in prostatic morphogenesis. Prostates exhibited distinctive lobe-specific growth and branching patterns in the ventral (VP) and lateral (LP) lobes, while exogenous FGF10 treatment shifted LP branching towards a VP branching pattern. Treatment with inhibitors of MEK1/2, Rho, Rho kinase, or MLCK significantly inhibited VP growth and blocked branching morphogenesis, further supporting critical roles for MEK/ERK and Rho/Rho kinase signaling pathways in prostatic growth and branching during development. We propose that MLCK-regulated MLC-P may be a central downstream target of both signaling pathways in regulating prostate morphogenesis.
    MeSH term(s) Animals ; Cell Differentiation ; Cell Proliferation ; Fibroblast Growth Factor 10/metabolism ; Gene Expression Regulation, Developmental ; MAP Kinase Signaling System ; Male ; Morphogenesis ; Organ Culture Techniques ; Prostate/growth & development ; Prostate/metabolism ; Rats ; rho-Associated Kinases/metabolism
    Chemical Substances Fgf10 protein, rat ; Fibroblast Growth Factor 10 ; rho-Associated Kinases (EC 2.7.11.1)
    Language English
    Publishing date 2021-12-04
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2701262-1
    ISSN 2218-273X ; 2218-273X
    ISSN (online) 2218-273X
    ISSN 2218-273X
    DOI 10.3390/biom11121829
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: Stem cells from a malignant rat prostate cell line generate prostate cancers

    Hu, Wen-Yang / Liu, Li-Feng / Afradiasbagharani, Parivash / Lu, Ran-Li / Chen, Zhen-Long / Hu, Dan-Ping / Birch, Lynn A / Prins, Gail S

    American journal of clinical and experimental urology

    2022  Volume 10, Issue 6, Page(s) 377–389

    Abstract: Cancer stem cells (CSCs) are resistant to conventional cancer therapies, permitting the repopulation of new tumor growth and driving disease progression. Models for testing prostate CSC-propagated tumor growth are presently limited yet necessary for ... ...

    Abstract Cancer stem cells (CSCs) are resistant to conventional cancer therapies, permitting the repopulation of new tumor growth and driving disease progression. Models for testing prostate CSC-propagated tumor growth are presently limited yet necessary for therapeutic advancement. Utilizing the congenic nontumorigenic NRP152 and tumorigenic NRP154 rat prostate epithelial cell lines, the present study investigated the self-renewal, differentiation, and regenerative abilities of prostate stem/progenitor cells and developed a CSC-based PCa model. NRP154 cells expressed reduced levels of tumor suppressor caveolin-1 and increased p-Src as compared to NRP152 cells. Gene knockdown of caveolin-1 in NRP152 cells upregulated p-Src, implicating their role as potential oncogenic mediators in NRP154 cells. A FACS-based Hoechst exclusion assay revealed a side population of stem-like cells (0.1%) in both NRP152 and NRP154 cell lines. Using a 3D Matrigel culture system, stem cells from both cell lines established prostaspheres at a 0.1% efficiency through asymmetric self-renewal and rapid proliferation of daughter progenitor cells. Spheres derived from both cell lines contained CD117
    Language English
    Publishing date 2022-12-25
    Publishing country United States
    Document type Journal Article
    ISSN 2330-1910
    ISSN 2330-1910
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Therapeutic Strategies for Restoring Perturbed Corneal Epithelial Homeostasis in Limbal Stem Cell Deficiency: Current Trends and Future Directions.

    Masood, Faisal / Chang, Jin-Hong / Akbar, Anosh / Song, Amy / Hu, Wen-Yang / Azar, Dimitri T / Rosenblatt, Mark I

    Cells

    2022  Volume 11, Issue 20

    Abstract: Limbal stem cells constitute an important cell population required for regeneration of the corneal epithelium. If insults to limbal stem cells or their niche are sufficiently severe, a disease known as limbal stem cell deficiency occurs. In the absence ... ...

    Abstract Limbal stem cells constitute an important cell population required for regeneration of the corneal epithelium. If insults to limbal stem cells or their niche are sufficiently severe, a disease known as limbal stem cell deficiency occurs. In the absence of functioning limbal stem cells, vision-compromising conjunctivalization of the corneal epithelium occurs, leading to opacification, inflammation, neovascularization, and chronic scarring. Limbal stem cell transplantation is the standard treatment for unilateral cases of limbal stem cell deficiency, but bilateral cases require allogeneic transplantation. Herein we review the current therapeutic utilization of limbal stem cells. We also describe several limbal stem cell markers that impact their phenotype and function and discuss the possibility of modulating limbal stem cells and other sources of stem cells to facilitate the development of novel therapeutic interventions. We finally consider several hurdles for widespread adoption of these proposed methodologies and discuss how they can be overcome to realize vision-restoring interventions.
    MeSH term(s) Humans ; Limbus Corneae ; Corneal Diseases/therapy ; Cornea ; Stem Cells ; Homeostasis
    Language English
    Publishing date 2022-10-16
    Publishing country Switzerland
    Document type Journal Article ; Review ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ZDB-ID 2661518-6
    ISSN 2073-4409 ; 2073-4409
    ISSN (online) 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells11203247
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Development and application of a low-priced duplex quantitative PCR assay based on SYBR Green I for the simultaneous detection of porcine deltacoronavirus and porcine sapelovirus.

    Lu, Si-Jia / Ma, Meng-Yao / Yan, Xiao-Guang / Zhao, Fu-Jie / Hu, Wen-Yang / Ding, Qing-Wen / Ren, Hao-Jie / Xiang, Yu-Qiang / Zheng, Lan-Lan

    Veterinarni medicina

    2023  Volume 68, Issue 3, Page(s) 106–115

    Abstract: Porcine deltacoronavirus (PDCoV) and porcine sapelovirus (PSV) are two viruses that can cause diarrhoea in pigs and bring great economic loss to the pig industry. In this research, a duplex real-time quantitative polymerase chain reaction (qPCR) assay ... ...

    Abstract Porcine deltacoronavirus (PDCoV) and porcine sapelovirus (PSV) are two viruses that can cause diarrhoea in pigs and bring great economic loss to the pig industry. In this research, a duplex real-time quantitative polymerase chain reaction (qPCR) assay based on SYBR Green I was developed to simultaneously detect PDCoV and PSV. No specific melting peaks were found in other porcine diarrhoea-associated viruses, indicating that the method developed in this study had good specificity. The detection limits of PDCoV and PSV were 1.0 × 10
    Language English
    Publishing date 2023-03-23
    Publishing country Czech Republic
    Document type Journal Article
    ZDB-ID 412227-6
    ISSN 0375-8427 ; 0590-5214
    ISSN 0375-8427 ; 0590-5214
    DOI 10.17221/79/2022-VETMED
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article: WNT2 is necessary for normal prostate gland cyto-differentiation and modulates prostate growth in an FGF10 dependent manner.

    Madueke, Ikenna C / Hu, Wen-Yang / Huang, Liwei / Prins, Gail S

    American journal of clinical and experimental urology

    2018  Volume 6, Issue 4, Page(s) 154–163

    Abstract: Wnt proteins are highly conserved secreted morphogens that function in organ development across species. This study investigates the role(s) of Wnt2 during prostate gland development. ...

    Abstract Wnt proteins are highly conserved secreted morphogens that function in organ development across species. This study investigates the role(s) of Wnt2 during prostate gland development.
    Language English
    Publishing date 2018-08-20
    Publishing country United States
    Document type Journal Article
    ISSN 2330-1910
    ISSN 2330-1910
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article: Isolation of stem-like cells from 3-dimensional spheroid cultures

    Hu, Wen-Yang / Hu, Dan-Ping / Xie, Lishi / Birch, Lynn A / Prins, Gail S

    Journal of visualized experiments. 2019 Dec. 13, , no. 154

    2019  

    Abstract: Despite advances in adult stem cell research, identification and isolation of stem cells from tissue specimens remains a major challenge. While resident stem cells are relatively quiescent with niche restraints in adult tissues, they enter the cell cycle ...

    Abstract Despite advances in adult stem cell research, identification and isolation of stem cells from tissue specimens remains a major challenge. While resident stem cells are relatively quiescent with niche restraints in adult tissues, they enter the cell cycle in anchor-free three-dimensional (3D) culture and undergo both symmetric and asymmetric cell division, giving rise to both stem and progenitor cells. The latter proliferate rapidly and are the major cell population at various stages of lineage commitment, forming heterogeneous spheroids. Using primary normal human prostate epithelial cells (HPrEC), a spheroid-based, label-retention assay was developed that permits the identification and functional isolation of the spheroid-initiating stem cells at a single cell resolution. HPrEC or cell lines are two-dimensionally (2D) cultured with BrdU for 10 days to permit its incorporation into the DNA of all dividing cells, including self-renewing stem cells. Wash out commences upon transfer to the 3D culture for 5 days, during which stem cells self-renew through asymmetric division and initiate spheroid formation. While relatively quiescent daughter stem cells retain BrdU-labeled parental DNA, the daughter progenitors rapidly proliferate, losing the BrdU label. BrdU can be substituted with CFSE or Far Red pro-dyes, which permit live stem cell isolation by FACS. Stem cell characteristics are confirmed by in vitro spheroid formation, in vivo tissue regeneration assays, and by documenting their symmetric/asymmetric cell divisions. The isolated label-retaining stem cells can be rigorously interrogated by downstream molecular and biologic studies, including RNA-seq, ChIP-seq, single cell capture, metabolic activity, proteome profiling, immunocytochemistry, organoid formation, and in vivo tissue regeneration. Importantly, this marker-free functional stem cell isolation approach identifies stem-like cells from fresh cancer specimens and cancer cell lines from multiple organs, suggesting wide applicability. It can be used to identify cancer stem-like cell biomarkers, screen pharmaceuticals targeting cancer stem-like cells, and discover novel therapeutic targets in cancers.
    Keywords DNA ; adult stem cells ; adults ; biochemical pathways ; biomarkers ; cell division ; cell lines ; chromatin immunoprecipitation ; drugs ; epithelial cells ; humans ; immunocytochemistry ; neoplasm cells ; neoplasms ; organoids ; proteome ; sequence analysis ; therapeutics ; tissue repair
    Language English
    Dates of publication 2019-1213
    Size p. e60357.
    Publishing place Journal of Visualized Experiments
    Document type Article
    ZDB-ID 2259946-0
    ISSN 1940-087X
    ISSN 1940-087X
    DOI 10.3791/60357
    Database NAL-Catalogue (AGRICOLA)

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  8. Article ; Online: Keratin Profiling by Single-Cell RNA-Sequencing Identifies Human Prostate Stem Cell Lineage Hierarchy and Cancer Stem-Like Cells.

    Hu, Wen-Yang / Hu, Dan-Ping / Xie, Lishi / Nonn, Larisa / Lu, Ranli / Abern, Michael / Shioda, Toshihiro / Prins, Gail S

    International journal of molecular sciences

    2021  Volume 22, Issue 15

    Abstract: Single prostate stem cells can generate stem and progenitor cells to form prostaspheres in 3D culture. Using a prostasphere-based label retention assay, we recently identified keratin 13 ( ...

    Abstract Single prostate stem cells can generate stem and progenitor cells to form prostaspheres in 3D culture. Using a prostasphere-based label retention assay, we recently identified keratin 13 (
    MeSH term(s) Adult ; Cells, Cultured ; Humans ; Keratins/metabolism ; Male ; Neoplastic Stem Cells/metabolism ; Neoplastic Stem Cells/pathology ; Primary Cell Culture ; Prostatic Neoplasms/metabolism ; Prostatic Neoplasms/pathology ; RNA/metabolism ; Single-Cell Analysis ; Young Adult
    Chemical Substances RNA (63231-63-0) ; Keratins (68238-35-7)
    Language English
    Publishing date 2021-07-28
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms22158109
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Isolation of Stem-like Cells from 3-Dimensional Spheroid Cultures.

    Hu, Wen-Yang / Hu, Dan-Ping / Xie, Lishi / Birch, Lynn A / Prins, Gail S

    Journal of visualized experiments : JoVE

    2019  , Issue 154

    Abstract: Despite advances in adult stem cell research, identification and isolation of stem cells from tissue specimens remains a major challenge. While resident stem cells are relatively quiescent with niche restraints in adult tissues, they enter the cell cycle ...

    Abstract Despite advances in adult stem cell research, identification and isolation of stem cells from tissue specimens remains a major challenge. While resident stem cells are relatively quiescent with niche restraints in adult tissues, they enter the cell cycle in anchor-free three-dimensional (3D) culture and undergo both symmetric and asymmetric cell division, giving rise to both stem and progenitor cells. The latter proliferate rapidly and are the major cell population at various stages of lineage commitment, forming heterogeneous spheroids. Using primary normal human prostate epithelial cells (HPrEC), a spheroid-based, label-retention assay was developed that permits the identification and functional isolation of the spheroid-initiating stem cells at a single cell resolution. HPrEC or cell lines are two-dimensionally (2D) cultured with BrdU for 10 days to permit its incorporation into the DNA of all dividing cells, including self-renewing stem cells. Wash out commences upon transfer to the 3D culture for 5 days, during which stem cells self-renew through asymmetric division and initiate spheroid formation. While relatively quiescent daughter stem cells retain BrdU-labeled parental DNA, the daughter progenitors rapidly proliferate, losing the BrdU label. BrdU can be substituted with CFSE or Far Red pro-dyes, which permit live stem cell isolation by FACS. Stem cell characteristics are confirmed by in vitro spheroid formation, in vivo tissue regeneration assays, and by documenting their symmetric/asymmetric cell divisions. The isolated label-retaining stem cells can be rigorously interrogated by downstream molecular and biologic studies, including RNA-seq, ChIP-seq, single cell capture, metabolic activity, proteome profiling, immunocytochemistry, organoid formation, and in vivo tissue regeneration. Importantly, this marker-free functional stem cell isolation approach identifies stem-like cells from fresh cancer specimens and cancer cell lines from multiple organs, suggesting wide applicability. It can be used to identify cancer stem-like cell biomarkers, screen pharmaceuticals targeting cancer stem-like cells, and discover novel therapeutic targets in cancers.
    MeSH term(s) Bromodeoxyuridine ; Cell Count ; Cell Cycle/physiology ; Cell Division ; Cell Separation/methods ; Cells, Cultured ; Flow Cytometry ; Humans ; Male ; Prostate/cytology ; Spheroids, Cellular ; Stem Cells/cytology
    Chemical Substances Bromodeoxyuridine (G34N38R2N1)
    Language English
    Publishing date 2019-12-13
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Video-Audio Media
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/60357
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Effects of Inorganic Arsenic on Human Prostate Stem-Progenitor Cell Transformation, Autophagic Flux Blockade, and NRF2 Pathway Activation.

    Xie, Lishi / Hu, Wen-Yang / Hu, Dan-Ping / Shi, Guangbin / Li, Ye / Yang, Jianfu / Prins, Gail S

    Environmental health perspectives

    2020  Volume 128, Issue 6, Page(s) 67008

    Abstract: Background: Inorganic arsenic (iAs) is an environmental toxicant associated with an increased risk of prostate cancer in chronically exposed populations worldwide. However, the biological mechanisms underlying iAs-induced prostate carcinogenesis remain ... ...

    Abstract Background: Inorganic arsenic (iAs) is an environmental toxicant associated with an increased risk of prostate cancer in chronically exposed populations worldwide. However, the biological mechanisms underlying iAs-induced prostate carcinogenesis remain unclear.
    Objectives: We studied how iAs affects normal human prostate stem-progenitor cells (PrSPCs) and drives transformation and interrogated the molecular mechanisms involved.
    Methods: PrSPCs were enriched by spheroid culture from normal human primary or immortalized prostate epithelial cells, and their differentiation capability was evaluated by organoid culture. Microarray analysis was conducted to identify iAs-dysregulated genes, and lentiviral infection was used for stable manipulation of identified genes. Soft agar colony growth assays were applied to examine iAs-induced transformation. For
    Results: Low-dose iAs (
    Conclusions: Low-dose iAs increased self-renewal and decreased differentiation of human PrSPCs by activating the p62-NRF2 axis, resulting in epithelial cell transformation. NRF2 is activated by iAs through specific autophagic flux blockade in progenitor cells, which may have potential therapeutic implications. https://doi.org/10.1289/EHP6471.
    MeSH term(s) Animals ; Arsenic/toxicity ; Cell Line ; Cell Transformation, Neoplastic/chemically induced ; Hazardous Substances/toxicity ; Humans ; Male ; Mice ; Mice, Nude ; NF-E2-Related Factor 2 ; Prostate ; Rats ; Stem Cells
    Chemical Substances Hazardous Substances ; NF-E2-Related Factor 2 ; Arsenic (N712M78A8G)
    Language English
    Publishing date 2020-06-11
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 195189-0
    ISSN 1552-9924 ; 0091-6765 ; 1078-0475
    ISSN (online) 1552-9924
    ISSN 0091-6765 ; 1078-0475
    DOI 10.1289/EHP6471
    Database MEDical Literature Analysis and Retrieval System OnLINE

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