Development of Small Molecule Inhibitor of PELP1 for Treating Advanced Breast Cancer

项目摘要/摘要：乳腺癌(BC)是最常见和第二致命的癌症 女性中的恶性肿瘤。治疗难治性BCA患者的新的有效治疗方法的开发 BC)和三重阴性BCA(TNBC)仍然是改善生存结果的最高需求。脯氨酸-， 谷氨酸和富亮氨酸蛋白1(PELP1)是一种原癌基因，在多发性核病变中起关键作用 受体(NR)的功能导致BC进展。PELP1在BCA中表达上调，促进 通过组蛋白修饰的表观遗传学改变，是预测BCA存活率差的指标，并有助于 药物治疗--耐药和转移。我的F31学位论文研究的重点是开发一个第一- 靶向致癌基因PELP1的一类小分子抑制剂(SMIPs)及其作为高级药物的疗效 BC疗法。我假设PELP1将NRs与表观遗传修饰物偶联，并通过SMIP靶向这个轴 在治疗tr-bc和tnbc方面都有疗效。我的初步数据显示铅化合物， SMIP34可抑制已建立的BCA细胞系的增殖，其IC50值在3-10μM之间。 提示SMIP34有能力阻断PELP1的致癌功能，包括降低细胞存活率和 侵袭性和促进细胞凋亡。目标1将定义SMIP34作为 PELP1抑制剂。我将使用多种生物物理方法来证实SMIP34与PELP1的直接相互作用。 我将使用有条件的CRISPR敲除PELP1来进一步确认特异性。我会评估他们的能力 SMIP34抑制PELP1癌基因相互作用组并评估下游信号通路的调节 使用全基因组方法，包括芯片序列和质谱分析。SMIP34治疗的疗效观察 对基因表达的影响将通过RNA-Seq进行测定和进一步分析，以确定潜在的相关 生物标志物。目的2将评估SMIP 34在治疗晚期乳腺癌中的有效性，包括临床前和 PDX型号。具有免疫能力的CD1小鼠模型将被用于研究毒性和确定 SMIP34的最大耐受量。患者来源的外植体(PDEX)模型将被用来描述 SMIP34对人BC瘤外植体生长的影响Tr-bc和tnbc同源模型，在 除了PDX原位异种移植模型外，还将用于测试治疗对肿瘤体积的影响， 疾病进展、最大耐受量和可观察到的毒性。我会用IHC分析肿瘤组织 确定其作用机制和疗效。我的长期职业目标是阐明 乳腺癌和妇科癌症中癌基因信号转导作用的研究 治疗。最终，F31奖学金将为我提供多种发展职业生涯的机会 独立的壁外资助的研究人员专注于女性癌症的新翻译进展 治疗学。

PROJECT SUMMARY/ABSTRACT: Breast cancer (BC) is the most prevalent and second most lethal malignancy in females. Development of novel effective therapies for patients with therapy-resistant BCa (TR- BC) and triple negative BCa (TNBC) remains the highest unmet need in improving survival outcomes. Proline-, glutamic acid, and leucine-rich protein 1 (PELP1), is a proto-oncogene that plays a critical role in multiple nuclear receptor (NR) functions leading to BC progression. PELP1 expression is upregulated in BCa, promotes epigenetic changes through histone modification, is a prognostic indicator of poor BCa survival, and contributes to drug therapy-resistance and metastases. The focus of my F31 dissertation research is to develop a First-in- Class small molecule inhibitor targeting oncogenic PELP1 (SMIPs) and elucidate the efficacy as an advanced BC therapy. I hypothesize that PELP1 couples NRs with epigenetic modifiers and targeting this axis with SMIPs will have therapeutic utility in treating both TR-BC and TNBC. My preliminary data indicates lead compound, SMIP34, inhibits proliferation in an established panel of BCa cell lines with an IC50 between 3-10μM. Results indicate that SMIP34 has ability to block PELP1 oncogenic functions including reduction of cell viability and invasiveness and promotion of apoptosis. Aim 1 will define the mechanisms by which SMIP34 functions as a PELP1 inhibitor. I will use multiple biophysical methods to confirm the direct interaction of SMIP34 with PELP1. I will further confirm specificity using conditional CRISPR knockout of PELP1. I will evaluate the ability of SMIP34 to inhibit the PELP1 oncogenic interactome and assess modulation of downstream signaling pathways using genome wide approaches including ChIP-Seq and Mass Spec analyses. The effects of SMIP34 treatment on gene expression will be determined by RNA-Seq and further analyzed in order to identify potential correlative biomarkers. Aim 2 will evaluate the utility of SMIP 34 in treating advanced breast cancer using preclinical and PDX models. Immuno-competent CD1 mouse models will be used to study toxicity and to determine the maximum tolerated dose of SMIP34. Patient derived explant (PDEX) models will be utilized to characterize the ex vivo effects of SMIP34 on growth of human BC tumor explants. Both TR-BC and TNBC syngenic models, in addition to PDX orthotopic xenograft models, will be utilized to test the effects of treatment on tumor volume, disease progression, maximum tolerated dose, and observable toxicity. I will use IHC analyses of tumor tissues to confirm mechanism of action and efficacy. My long-term career objective is elucidating the mechanistic contributions oncogenic signaling in breast and gynecological cancers for the purpose of developing targeted therapies. Ultimately, the F31 Fellowship will provide multiple opportunities to develop my career as an independent extramurally funded investigator focused on novel translational advances in women's cancer therapeutics.