Mitochondria specific metabolomic signature in triple negative breast cancer meta

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。代谢物是蛋白质活性的最终产物，不那么复杂，更稳定，很容易在临床环境中进行测量。生长活跃的细胞(肿瘤)的大部分新陈代谢发生在细胞质和线粒体。后者被认为是细胞的能量屋，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。通过融合含有感兴趣线粒体的去核细胞和Rho0受体细胞(含有去除的线粒体的细胞)来构建胞质杂交体 DNA)。Kaipparettu博士在体外和活体环境中使用这项技术，证明了TN BCA的线粒体逆行调节(MRR)进程中的关键作用。在这里，我们建议将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结合起来，以评估一个具有临床挑战性的问题，“我们如何更好地区分转移性TN BCA？”从战略上讲，我们将使用Cybrid系统分析转移性BCA中线粒体的特异性变化。Kaipparettu实验室已经提供了来自不同TN乳房来源的非癌和癌核背景的Rho0细胞。我们将TN良性乳腺上皮的线粒体移植到这些受体细胞中，这些良性乳腺上皮非到中度转移和高度转移的BCA。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，将使用气/液耦合质谱仪和代谢表型相结合的方法来描述线虫和肿瘤的代谢组。代谢谱将使用已建立的生物统计和生物信息学管道与生物统计学家Creighton博士一起进行检查，以生成与TN BCA转移相关的代谢特征和途径。这些数据将与现有的来自患者来源的异种移植的患者来源的代谢图谱和具有长期临床随访的独立患者样本进行比较，以命名临床相关的代谢物。被提名的途径将使用细胞系和异种移植疾病模型来评估它们在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针对TN乳腺癌进展和转移的代谢组。在临床上，我们希望将这些转化为确定治疗TN乳腺癌的新药物靶点。

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. 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.