Regulation of the miRNA-directed Reprogramming of Normal Omental Fibroblasts

描述(申请人提供)：手术和积极化疗在卵巢癌(OvCa)中有20%的成功率，这意味着迫切需要新的治疗方法。卵巢肿瘤由7%-83%的间质组成，它推动肿瘤进展，增加血管生成，促进转移，但目前的治疗仅针对癌细胞1，2。虽然肿瘤间质由多种细胞类型组成，但癌症相关成纤维细胞(CAF)是主要成分。Lengyel和Peter实验室最近报道，与正常或癌旁成纤维细胞相比，在卵巢CAF中，miR-31和miR-214的微RNA(MiRNA)miR-31和miR-214下调，miR-155上调。此外，研究表明，在共培养过程中，OvCa细胞通过下调miR-214和miR-31，上调miR-155，足以将正常的大眼成纤维细胞(NOF)重新编程为CAF。通过转染miRNAs和miRNA抑制剂来模拟这种放松调控，可以诱导正常成纤维细胞向CAF的功能性转化，其定义是在体外和体内都增加了成纤维细胞的运动性和促进了肿瘤的进展。此外，逆转实验导致CAF逆转为正常成纤维细胞3。MiRNA重编程的正常成纤维细胞和患者来源的CAF共享大量高度上调的趋化因子基因，这些基因对CAF功能3至关重要。最高表达的趋化因子CCL5被发现是miR-214 3的直接靶标。这些结果表明，在卵巢癌进展过程中，NOF通过调节miRNA的表达而被重新编程为CAF。然而，OvCa通过这些miRNAs调节NOF到CAF重编程的因素和机制尚不清楚。因此，我建议确定在NOF到CAF重编程过程中调节miR-155、miR-214和miR-31表达的癌症衍生因子和成纤维细胞信号通路，这将导致识别卵巢癌用于重编程肿瘤间质的关键因素和机制。

DESCRIPTION (provided by applicant): Surgery and aggressive chemotherapy has a 20% success rate in ovarian cancer (OvCa) meaning new therapies are desperately needed. Ovarian tumors are composed of 7-83% stroma, which drives tumor progression, increases angiogenesis, and promotes metastasis, yet current therapies are aimed at targeting the cancer cells alone 1,2. While the tumor stroma consists of a number of cell types, cancer associated fibroblasts (CAFs) are a major constituent. The Lengyel and Peter labs recently reported that in ovarian CAFs, the micro-RNAs (miRNA) miR-31 and miR-214 are downregulated while miR-155 is upregulated when compared to normal or tumor-adjacent fibroblasts 3. Additionally, it was shown that OvCa cells are sufficient to reprogram normal omental fibroblasts (NOFs) into CAFs through downregulation of miR-214, miR-31, and upregulation of miR-155 during coculture. Mimicking this deregulation by transfecting miRNAs and miRNA inhibitors induced a functional conversion of normal fibroblasts into CAFs as defined by increased fibroblast motility and promotion of tumor progression both in vitro and in vivo. Furthermore, the reverse experiment resulted in the reversion of CAFs into normal fibroblasts3. The miRNA-reprogrammed normal fibroblasts and patient-derived CAFs shared a large number of upregulated genes highly enriched in chemokines, which are known to be important for CAF function 3. The most highly upregulated chemokine, CCL5, was found to be a direct target of miR- 214 3. These results indicate that during the process of ovarian cancer progression, NOFs are reprogrammed to become CAFs through the regulation of miRNA expression. However, the factors and the mechanisms in which OvCa regulates NOF to CAF reprogramming through these miRNAs are not understood. Therefore I propose to determine the cancer-derived factors and the fibroblast signaling pathways that regulate the expression of miR-155, miR-214, and miR-31 during NOF to CAF reprogramming, which would result in the identification of key factors and mechanisms used by ovarian cancer to reprogram the tumor stroma.