Mitochondria specific metabolomic signature in triple negative breast cancer meta

描述（由申请人提供）：对于三阴性BCa（TN BCa），目前缺乏对驱动途径的了解，因此通常使用更通用的疗法进行治疗。目前还没有临床上可接受的治疗TN BCa的目标，并预测其转移的潜力。代谢产物是蛋白质活性的终产物，并且不太复杂，更稳定，并且可以在临床环境中容易地测量。在活跃生长的细胞（肿瘤）中，大部分代谢发生在细胞质和线粒体中。后者被认为是细胞的能量屋，与肿瘤的发展和进展密切相关，因此是化疗的潜在靶点。随着跨线粒体胞质杂交体（cybrid）技术的出现，现在可以在确定的核背景下检查肿瘤相关线粒体对肿瘤生长和发育的具体贡献。胞质杂交体通过将含有感兴趣的线粒体的去核细胞与rho 0受体细胞（含有消融的线粒体的细胞）融合来构建。 DNA）。在体外和体内环境中使用这种技术，Kaipparettu博士已经证明了TN BCa的线粒体逆行调节（MRR）进展的关键作用。在这里，我们建议将联合收割机Kaipparettu博士（PI）在BCa和cybrid技术方面的专业知识与Sreekumar博士（Co-I）在BCa代谢组学方面的专业知识相结合，以评估一个具有临床挑战性的问题，“我们如何才能更好地区分转移性TN BCa？“从战略上讲，我们将使用cybrid系统来分析转移性BCa中的线粒体特异性改变。来自不同TN乳腺来源的非癌和癌核背景的rho 0细胞已经在Kaipparettu实验室中可用。我们将从TN良性乳腺上皮细胞非中度转移性和高度转移性BCa中移植线粒体到这些受体细胞中。将通过下一代测序确认由此产生的胞质杂交体的线粒体功能、核起源，并使用体外和体内肿瘤形成/侵袭测定评价表型变化。在此之后，将使用气/液色谱-联用质谱和代谢表型分析的组合来分析胞质杂交体和肿瘤的代谢组。将使用已建立的生物统计学和生物信息学管道与生物统计学家Creighton博士一起检查代谢谱，以生成与TN BCa转移相关的代谢特征和途径。将数据与来自患者来源异种移植物和独立患者样本的现有患者来源代谢谱进行比较，以确定临床相关代谢物。将使用该疾病的细胞系和异种移植模型评价指定途径在TN BCa进展中的作用。总的来说，我们希望开发出第一种用于TN乳腺癌进展和转移的α驱动代谢组。在临床上，我们希望这些被翻译为确定新的药物靶点的TN乳腺癌。

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. Metabolites are the end products of protein activity and are less complex, more stable, and can be easily measured in a clinical setting. Most of the metabolism in an actively growing cell (tumor) occurs in the cytoplasm and mitochondria. The latter 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 rho0 recipient cells (cells harboring ablated mitochondrial DNA). Using this technology in both an in vitro and in vivo setting, Dr. Kaipparettu has demonstrated a key role for mitochondrial retrograde regulation (MRR) progression of TN BCa. Here we propose to combine Dr. Kaipparettu's (PI) expertise in BCa and cybrid technology with Dr. Sreekumar's (Co-I) expertise on BCa metabolomics to evaluate a clinically challenging question, "How can we better distinguish metastatic TN BCa?" Strategically, we will use cybrid system to analyze the mitochondria specific alterations in metastatic BCa. rho0 cells from different TN breast-derived non-cancerous and cancerous nuclear background are already available in Kaipparettu lab. Into these recipient cells, we will transplant the mitochondria from TN benign breast epithelium non-to-moderately metastatic and highly metastatic BCa. 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, the cybrids and tumors will be profiled for their metabolome using a combination of gas/liquid chromatography-coupled mass spectrometry and metabolic phenotyping. The metabolic profiles will be examined using an established biostatistics and bioinformatics pipelines together with the biostatistician Dr. Creighton to generate metabolic signatures and pathways associated with TN BCa metastasis. The data will be compared with existing patient-derived metabolic profiles from patient-derived xenografts and independent patient specimens having long term clinical follow up to nominate clinically relevant metabolites. The nominated pathways will be evaluated for their role in TN BCa progression using cell line and xenograft models of the disease. Overall, we expect to develop the first-of-its-kind mitochondria-driven metabolome for TN breast cancer progression and metastasis. Clinically, we expect these to be translated to identify new drug targets for TN breast cancer.