GENETIC EVENTS IN HUMAN PROSTATE CELL TRANSFORMATION

描述（申请人的描述）：本提案的目标是 a）为以下人员提供结构化的分子遗传学培训计划： 申请人发展成为一个独立的研究人员，和B）研究 与克服衰老有关的遗传事件（即永生化） 在人类前列腺上皮细胞中。 许多文献研究 前列腺癌中基因组丢失和获得的详细模式。 但是，在这方面， 这些遗传事件或组合的功能意义 基因事件的发生还不清楚。 克服衰老可以考虑 肿瘤发生中的一个关键事件，具有明确的遗传基础。 在 提出的模型，表达病毒的人前列腺上皮培养物（HPEC）， 癌蛋白缺乏正常的pRB和/或p53调节功能（基因通常 在临床前列腺癌中改变）并且具有延长的寿命。 这些 体外事件使这些细胞处于额外的“自发” 与永生化相关的遗传和表观遗传改变。 的 申请人将测试这些基因的组合的假设， 事件，补充p53和/或pRb的损失，是重要的克服 体外前列腺癌中的细胞衰老。 此外，他们建议 这些途径在临床前列腺癌标本中发现。 他们的 具体目标包括：I）建立和表征体外模型 具有永生HPEC的系统，所述永生HPEC通过以下方式功能性地丧失p53和/或Rb： 选择性HPV 16 E6和/或E7逆转录病毒感染，ii）鉴定 额外的遗传和表观遗传事件，主要是事件的组合， 与克服p53-和Rb-连接途径的衰老相关，iii） 为了重新表达这些在永生化中丢失或获得的基因组区域， 永生和正常的HPEC，和iv）将这些体外事件与 已经丢失p53和/或Rb的临床前列腺癌样品 功能 这将为分子遗传学的研究提供新的视角 和与克服衰老相关的表观遗传事件，通过p53和/或 体外前列腺上皮细胞Rb功能丧失。 除了 为了解决这一在人类前列腺肿瘤形成中的关键机制作用， 将为申请人提供结构化的分子培训计划， 凯瑟琳·雷兹尼科夫博士实验室的遗传学研究

DESCRIPTION (Applicant's Description): The objectives of this proposal are to a) provide a structured, training program in molecular genetics for the applicant to develop into an independent researcher, and b) study the genetic events associated with overcoming senescence (i.e. immortalization) in human prostate epithelial cells. Numerous documentational studies have detailed patterns of genomic loss and gain in prostate cancers. However, the functional significance of many of these genetic events or combinations of genetic events remains unclear. Overcoming senescence can be considered a critical event in tumorigenesis that has a clear genetic basis. In the proposed model, human prostate epithelial cultures (HPECs) expressing viral oncoproteins lack normal pRB and/or p53 regulatory functions (genes commonly altered in clinical prostate cancers) and have an extended lifespan. These in vitro events place these cells at high risk for additional 'spontaneous' genetic and epigenetic alterations associated with immortalization. The applicants will test the hypothesis that combinations of these genetic events, complementing p53 and/or pRb loss, are important in overcoming cellular senescence in in vitro prostate cancer. In addition, they propose that these pathways are found in clinical prostate cancer specimens. Their SPECIFIC AIMS include: I) to establish and characterize an in vitro model system with immortal HPECs that have functionally lost p53 and/or Rb by selective HPV16 E6 and/or E7 retroviral infection, ii) to identify additional genetic and epigenetic events, chiefly combinations of events, associated with overcoming senescence for p53- and Rb-linked pathways, iii) to reexpress these genomic regions lost or gained at immortalization in immortal and normal HPECs, and iv) to correlate these in vitro events with clinical prostate cancer samples that have lost either p53 and/or Rb function. The proposal will provide new i n sight into molecular genetic and epigenetic events associated with overcoming senescence, via p53 and/or Rb loss of function, in prostate epithelial cells in vitro. In addition to addressing this critical mechanistic role in human prostate neoplasia, it will provide the applicant with a structured training program in molecular genetics in the laboratory of Dr. Catherine Reznikoff.