ZIA BC 005093 (ZIA) | |||
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Title | Growth-Differentiation Factors in Organogenesis | ||
Institution | NCI, Bethesda, MD | ||
Principal Investigator | Perantoni, Alan | NCI Program Director | N/A |
Cancer Activity | N/A | Division | CCR |
Funded Amount | $673,059 | Project Dates | 00/00/0000 - 00/00/0000 |
Fiscal Year | 2016 | Project Type | Intramural |
Research Topics w/ Percent Relevance | Cancer Types w/ Percent Relevance | ||
Cancer (100.0%) Chemotherapy (5.0%) Childhood Cancers (50.0%) |
Kidney Disease (90.0%) Urinary System (90.0%) Kidney Cancer (90.0%) Wilm's Tumor (90.0%) |
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Research Type | |||
Endogenous Factors in the Origin and Cause of Cancer Development and Characterization of Model Systems |
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Abstract | |||
The Renal Differentiation and Neoplasia Section studies inductive signaling in tissue development/morphogenesis and, in parallel, its dysregulation in tumorigenesis with emphasis on the ligands that mediate normal tissue interactions and the pathways and targets that are activated in response to signaling. Our focus has been on development of the urogenital tract, which features reciprocal interactions between two distinct mesodermal progenitors, highly coordinated tissue movements, mesenchymal-epithelial transition (MET), integration of structures from different lineages, reiterative cycles of development, and a tumor that caricatures nephrogenesis. More specifically we are interested in the signaling mechanisms that direct metanephric mesenchyme (MM) to convert to the epithelia of the nephron. Wilms tumor (WT) is characterized by an expanded blastemal/progenitor population with a restricted capacity for epithelial conversion (MET). It is our long-term goal to identify targets on which WT cells depend for survival or dysregulated signaling that can be reprogrammed to allow tumor cells to differentiate to a more benign phenotype. We have determined that the cytokine leukemia inhibitory factor (LIF) in combination with Rho kinase inhibitor (ROCKi) maintains and selectively expands the Six2+ nephronic stem cell population in culture. Moreover, these propagated stem cells retain their capacity to convert to all segments of the nephron, demonstrating that they are multipotent progenitors. LIF functions principally through activation of STATs 1, 3 and 5 and up regulates the expression of several renal stem cell markers, e.g., Six2 and Pax2. Mechanistically, we have now found that LIF stimulates JNK activation, which induces MM proliferation and enhances cell competence to differentiate. The Rho kinase inhibitor (ROCKi) attenuates the LIF-induced Jnk activation thus inhibiting the differentiation of the progenitor. An investigation into the mechanism(s) mediated by LIF/ROCKi in these cells revealed that our conditions facilitate the nuclear localization of Yes-associated protein (YAP), a transcriptional co-activator and component of the Hippo signaling pathway. Furthermore, silencing Yap gene expression by siRNA knockdown in MM cells decreased the expression of progenitor markers and increased levels of MET markers, suggesting that YAP maintains MM cells in an undifferentiated state. Since YAP interacts with Tead transcription factors, we also determined that canonical Yap signaling through Tead activation is required for YAP-dependent transcription and MM progenitor cell maintenance. Since last year, we have now reported optimized niche conditions for the propagation of mouse Six2+ nephronic progenitors. This involved treatment of FACs sorted GFP-Six2+ cells with a Wnt agonist and Bmp7 in addition to LIF/ROCKi. These cells could be maintained for several passages and induced to form all segments of the nephronic epithelia. This culture system of MM provides unique opportunities to comprehensively address key mechanisms involved in renal progenitor maintenance and differentiation and raises the possibility that they may be applied to models of tissue repair/regeneration. We have now also applied similar conditions to the propagation of human Wilms tumor cells and determined that these same factors selectively expand the Six2+ progenitor from tumor tissues. Given the belief that these cells provide the driving force for Wilms tumors, expansion of this population in culture may provide an important tool for developing individual treatments for Wilms tumor patients. In order to better understand the role that LIF-induced Stat signaling has in kidney development, we are evaluating Stat3 mutant mice, which develop kidneys that are dramatically reduced in size. During the course of our mouse genetic studies we discovered a significant role for Stat3 in bone development. We have determined that the conditional loss of Stat3 causes a phenot |