Probing Mediator 12 function in uterine fibroids

 描述(申请人提供)：子宫肌瘤，也被称为子宫肌瘤，是一种良性的纤维性平滑肌肿瘤，影响着令人震惊的75%-80%的女性一生，并构成世界范围内的主要健康问题。令人震惊的是，对它们的研究仍然不足，几乎没有非手术治疗选择。在该领域的一项重大突破中，被称为MED12的转录介质复合体亚单位12的突变在肌瘤中被发现的频率非常高，这意味着MED12突变是驱动突变的因素。然而，MED12及其突变是如何导致肌瘤的完全未知。我们实验室的长期目标是发现驱动肌瘤的新分子事件。这种R21探索性应用的直接目标是验证这样一种假设，即MED12突变通过在肌瘤中产生改变的MED12/中介细胞周期，导致细胞增殖/凋亡和促纤维化基因的异常表达而发挥作用。我们将确定正常子宫肌层和子宫肌瘤原代细胞中野生型和突变型MED12以及其他关键介质亚单位的全基因组结合谱。利用生物信息学和芯片定量聚合酶链式反应分析，我们将确定MED12是否主要通过靶向转录因子如核受体和Smad蛋白、介体复合体和RNA聚合酶II之间的通讯发挥作用，并确定这种相互作用在肌瘤中是缺陷/改变的。总之，这些结果将首次确定肌瘤MED12的分子缺陷，从而以前所未有的方式推进这一领域。此外，拟议的探索性分析将为这一高度未被研究的人类健康问题的未来RO1类型研究提议产生新的假设。这项拟议的工作具有科学、翻译和临床意义，因为我将为MED 12在肌瘤发展中的关键驱动机制提供一个新的视角，并为确定治疗这种疾病的其他治疗靶点和策略开辟可能性。它具有创新性，因为它代表了对以前未意识到的MED12在肌瘤病理生物学中的作用的第一次系统探索。

 DESCRIPTION (provided by applicant): Uterine leiomyomas, also known as uterine fibroids, are benign, fibrotic smooth muscle tumors that affect an alarming 75-80% of women in their lifetime and constitute a major health problem worldwide. They remain shockingly understudied with few nonsurgical therapeutic options. In a major breakthrough in the field, mutations in the transcriptional mediator complex subunit 12, termed Med 12, have been identified at very high frequency in leiomyoma implying that Med12 mutations are the driver mutations. However, how Med12 and its mutations contribute to leiomyoma is completely unknown. The long term goal of our laboratory is to discover novel molecular events driving leiomyoma. The immediate goal of this R21-exploratory application is to test the hypothesis that Med12 mutations function by generating an altered Med12/mediator cistrome in leiomyoma, leading to abnormal expression of cell proliferation/apoptosis and profibrotic genes. We will determine the genome wide binding profile of wild type and mutant Med12 and other key mediator subunits in normal myometrial and leiomyoma primary cells. Using bioinformatic and ChIP qPCR analysis we will determine whether Med12 functions primarily by targeting communication between transcription factors such as nuclear receptors and Smad proteins, mediator complex and RNA polymerase II and establish that such interactions are defective/altered in leiomyoma. Together, these results will determine for the first time molecular defects in Med12 in leiomyoma thereby advancing the field in an unprecedented manner. Additionally, the proposed exploratory analyses will generate new hypothesis for future RO1-type research proposals in this highly understudied human health problem. The proposed work is scientifically, translationally, and clinically significant because i will provide a new perspective on the key driver mechanisms of Med 12 function in leiomyoma development and open up possibilities for identifying additional therapeutic targets and strategies to treat this disease. It is innovative because it represents the first systematic exploration of previously unrealized roles of Med12 in leiomyoma patho-biology.