Mitochondria-nuclear crosstalk in triple negative breast cancer racial disparity

描述(申请人提供)：对于三阴性BCA(TN BCA)，目前对驾驶员路径缺乏了解，因此经常使用更通用的治疗方法。目前，对于TN BCA的治疗和预测其转移的可能性，还没有临床上可以接受的靶点。虽然BCA在高加索(CA)女性中的发病率较高，但非裔美国人(AA)女性因BCA而死亡的比例较高。重要的是，AA患者更有可能被诊断为三阴性(TN)BCA，并且比CA处于更晚期。线粒体被认为是细胞的能量库，与肿瘤的发生和发展密切相关，因此成为潜在的化疗靶点。随着跨线粒体胞质杂交(Cybrid)技术的出现，现在有可能在特定的核背景下研究肿瘤相关线粒体对肿瘤生长和发育的特定贡献。胞质杂交是通过融合含有感兴趣线粒体的去核细胞和？0受体细胞(含有去除的线粒体DNA的细胞)来构建的。在体外和体内使用这项技术，我们已经证明了线粒体逆行调节(MRR)在TN BCA进展中的关键作用。我们还确定了癌症线粒体对Src致癌途径翻译后修饰的关键调控。我们对AA和CA TN BCA细胞系线粒体的初步分析表明，在AA和CA细胞中，核基因的线粒体逆行调节(MRR)是不同的。在这里，我们建议将Kaipparettu博士在BCA和Cybrid技术方面的专业知识与Lewis博士在生成患者来源的异种移植物方面的专业知识和Creighton博士在生物信息学方面的专业知识结合起来，以评估一个具有临床挑战性的问题，“我们如何通过其MRR更好地区分AA和CA TN BCA？”策略上，我们将使用AA和CA TN BCA患者来源的异种移植物产生的Cybrid系统来分析转移性BCA中线粒体的特异性变化。？0来自不同TN乳房来源的AA和CA核背景的细胞已经在Kaipparettu实验室获得。由此产生的胞质将通过下一代测序确认其线粒体功能和核来源，并使用体外和体内肿瘤形成/侵袭试验评估表型变化。在此之后，带有AA和CA线粒体的胞质杂交体将被用于其Src途径的修饰。为了确定新的途径，将通过微阵列分析线虫的MRR，并将使用已建立的生物统计和生物信息学管道(Oncomine概念图)与生物统计学家Creighton博士一起检查基因表达，以生成与AA和CA TN BCA转移相关的致癌特征和途径。被提名的通路将使用细胞系和异种移植模型来评估它们在TN BCA进展中的作用。总体而言，我们希望开发首个线粒体驱动的AA和CA TN BCA进展的致癌信号。临床上，我们希望将这些转化为AA、TN、BCA的新药靶点。

DESCRIPTION (provided by applicant): For Triple Negative BCa (TN BCa), there is a current lack of understanding of driver pathways and hence are often treated using more generic therapies. Currently there are no clinically accepted targets for the treatment for TN BCa and to predict its potential to metastasize. while BCa incidence is higher in Caucasian (CA) women, death due to BCa is higher in African American (AA) women. Importantly, AA patients are more likely to be diagnosed with triple negative (TN) BCa and at a more advanced stage than CA. Mitochondria is considered the energy house of the cell and has been strongly implicated in tumor development and progression and thus serving as a potential target for chemotherapy. With the advent of Transmitochondrial cybrid (cybrid) technology, it is now possible to examine the specific contribution of tumor-associated mitochondria to neoplastic growth and development under a defined nuclear background. Cybrids are constructed by fusing enucleated cells harboring mitochondria of interest with ?0 recipient cells (cells harboring ablated mitochondrial DNA). Using this technology in both an in vitro and in vivo setting, we have demonstrated a key role for mitochondrial retrograde regulation (MRR) progression of TN BCa. We have also identified critical regulation of post-translational modification of Src oncogenic pathway by cancer mitochondria. Our preliminary analysis using cybrids with mitochondria from AA and CA TN BCa cell lines suggest that mitochondrial retrograde regulation (MRR) of nuclear genes are different in AA and CA cells. Here we propose to combine Dr. Kaipparettu's (PI) expertise in BCa and cybrid technology with Dr. Lewis expertize in generating patient derived xenografts and Dr. Creighton's expertise in bioinformatics to evaluate a clinically challenging question, "How can we better distinguish AA and CA TN BCa by its MRR?" Strategically, we will use cybrid system generated from AA and CA TN BCa patients-derived xenografts to analyze the mitochondria specific alterations in metastatic BCa. ?0 cells from different TN breast-derived AA and CA nuclear background are already available in Kaipparettu lab. The cybrids thus generated will be confirmed for their mitochondrial function, nuclear origin by next-gen sequencing and evaluated for phenotypic changes using in vitro and in vivo tumor forming/invasion assays. Following this, cybrids with AA and CA mitochondria will be profiled for their Src pathway modification. To identify novel pathways, the MRR of cybrids will be analyzed by microarray and the gene expression will be examined using an established biostatistics and bioinformatics pipelines (Oncomine Concept Mapping) together with the biostatistician Dr. Creighton to generate oncogenic signatures and pathways associated with AA and CA TN BCa metastasis. The nominated pathways will be evaluated for their role in TN BCa progression using cell line and xenograft models. Overall, we expect to develop the first- of-its-kind mitochondria-driven oncogenic signature for AA and CA TN BCa progression. Clinically, we expect these to be translated to identify new drug targets for AA TN BCa.