@article{Regeneration:2807,
      recid = {2807},
      author = {Moline, Daniel Clark},
      title = {Single Cell RNA Sequencing of Prostate Samples Yields  Candidate Factors Necessary for Prostate Regeneration},
      publisher = {University of Chicago},
      school = {Ph.D.},
      address = {2021-03},
      pages = {122},
      abstract = {      The human prostate is a significant source of  disease burden for adult males. In the malignant context,  prostate cancer is the most common non-cutaneous cancer in  males. Benign prostatic hyperplasia is also common,  affecting a majority of males over the age of 60. This high  disease burden has been a focus of study for many years,  but an answer to the ultimate question of what predisposes  the prostate to such a high level of disease has eluded  researchers. One possible explanation for the disease  burden of the prostate may lie in the capacity of the  prostate for hormone-dependent regeneration. This is a  process by which the prostate shrinks in the absence of  androgen signaling, and regenerates to its original size  when androgen is reintroduced to the system. The study of  hormone-dependent regeneration has allowed researchers to  analyze the progenitor cell populations of the prostate.	In  this thesis, single cell RNA-Seq approaches were used to  investigate the progenitor cell populations of the  prostate. First, I investigated cells harvested from  monolayer and organoid culture conditions to better  understand the progenitor populations present in these  models and differences between the models overall. This  yielded evidence that prostate progenitor cells expressing  Keratin 13 were preserved in both monolayer and organoid  conditions. This presence of prostate progenitor cells was  validated using immunofluorescence microscopy targeting  Keratin 13 protein. In comparing the single cell RNA-Seq  data from the two culture conditions, we were able to  observe an enrichment of proliferating populations in the  monolayer sample and an enrichment of intermediate cells in  the organoid sample. Further comparison of these in vitro  samples with in vivo prostate data gathered by another lab  provided evidence that the in vitro samples were enriched  for proliferating cells while the in vivo sample contained  terminally differentiated cell populations that were not  observed in vitro. These data provide an in-depth  validation of the preservation of prostate progenitor cells  in commonly used in vitro models, as well as providing  insights into the different cell populations selected for  in monolayer and organoid culture conditions respectively.  
	Application of single cell RNA-Seq approaches to in vivo  mouse prostate led to the identification of a luminal  progenitor cell population in both the intact and castrate  mouse prostate. These cells expressed luminal keratins as  well as multiple putative progenitor cell markers. The  presence of luminal progenitor cells in the mouse prostate  was also validated using both immunofluorescence microscopy  and flow cytometry. Pathway analysis of the expression data  from luminal progenitor cells allowed for the selection of  candidate factors likely to contribute to the prostate  progenitor cell phenotype. Small molecule inhibitor  treatment targeting two of these factors, Yap1 and Bcl-2,  caused a significant decrease in in vitro regeneration of  organoids derived from both mouse and human cells. These  results provide in-depth expression data for luminal  progenitor cells and also identify factors necessary for  the prostate progenitor cell phenotype. These factors can  be leveraged to better understand luminal progenitor cells  and possibly be used to treat prostate disease in the  future.},
      url = {http://knowledge.uchicago.edu/record/2807},
      doi = {https://doi.org/10.6082/uchicago.2807},
}