Regulation of Mitochondrial Metabolism by Tyr-phosphorylated ATP Synthase Alpha-Subunit and its Therapeutic Implications in Prostate Cancer

项目摘要 癌细胞线粒体改变其代谢表型以满足高能量需求和 大分子合成。肿瘤对致癌酪氨酸激酶信号的急性依赖性支持其 然而，快速增殖与支持这些高能的线粒体的激酶活性直接相关 流程仍然模糊不清。偶然的是，我们发现了一种非受体酪氨酸激酶，ACK1， 磷酸化三磷酸腺苷合成酶F1α亚单位(ATP5F1a)在Tyr243和Tyr246(成熟蛋白中的Tyr200和203， 分别)，这增加了ATP合成酶的活性，特别是在癌细胞。从机械上讲，ATP5F1a- 磷酸化不仅排除了它与其生理抑制物ATPase抑制因子1(IF1)的结合，而且 还产生了从结合的非催化核苷酸延伸到表面的支撑结构， 降低了柔韧性，从而增加了酶的稳定性。新型AcK1抑制剂(R)-9b 逆转这一过程，诱导有丝分裂，减缓肿瘤生长。始终如一地， ATP5F1a-磷酸化在正常前列腺向恶性进展的过程中被观察到。总的来说，这些 数据为癌细胞对Tyr-Kinase的沉迷而“线粒体成瘾”提供了分子证据 揭示(R)-9b是一种“有丝分裂原蛋白”，能满足癌细胞独特的代谢需求。 耐去势前列腺癌(CRPC)仍然是一种无法治愈的恶性肿瘤，治疗选择有限。 是全球男性死亡的一个重要原因(15)。新城疫疗效有限，耐药性快速发展 苯扎鲁胺和阿比拉通、AR拮抗剂的治疗已经建立了一个新的范式--实现现实 缓解后，其他癌症特异性途径，包括代谢，必须受到影响。这项建议是针对 AcK1/ATP5F1a线粒体代谢调控特性的详细研究 (R)-9b克服恩扎鲁胺和阿比特龙耐药CRPC的信号和检测能力。这个 具体目标如下： 具体目标1.研究ATP5F1a-磷酸化调节其在前列腺中活性的机制 癌症 特定目的2.探讨AcK1/ATP5F1a信号在前列腺癌模型中的作用 具体目的3.苯扎鲁胺和阿比特龙中AcK1/ATP5F1a信号转导的详细体内特征 小鼠前列腺癌模型和患者来源的异种移植(PDXs)的耐药性

Project Abstract Cancer cell mitochondria switch their metabolic phenotypes to meet the challenges of high-energy demand and macromolecular synthesis. Acute dependence of tumors on oncogenic tyrosine kinase signaling support their rapid proliferation, however, direct relevance of the kinase activity for mitochondria to support these high-energy processes remains obscure. Serendipitously, we uncovered that a non-receptor tyrosine kinase, ACK1, phosphorylates ATP synthase F1 α-subunit (ATP5F1a) at Tyr243 and Tyr246 (Tyr200 & 203 in mature protein, respectively) that increased ATP synthase activity specifically in the cancer cells. Mechanistically, ATP5F1a- phosphorylation not only excluded its binding to its physiological inhibitor, ATPase Inhibitory Factor 1 (IF1), but also created a supporting structure that extended from the bound non-catalytic nucleotide to the surface, reducing the flexibility and thereby increasing the stability of the enzyme. A new class of ACK1 inhibitor, (R)-9b reversed this process, inducing mitophagy and mitigating tumor growth. Consistently, a marked increase in ATP5F1a-phosphorylations was observed as normal prostate progressed to the malignant stage. Overall, these data provide the molecular evidence for cancer cell `mitochondrial addiction’ to Tyr-kinase indulgence, and reveals (R)-9b as a ‘mitocan’ that compromises the unique metabolic needs of cancer cells. Castration resistant prostate cancer (CRPC) remains an incurable malignancy with limited treatment options and is a significant cause of deaths in men worldwide (15). Limited efficacy and rapid development of resistance for Enzalutamide and Abiraterone, AR antagonist treatment have established a new paradigm-to achieve realistic remission, other cancer specific pathways, including metabolic must be compromised. This proposal is directed towards detailed characterization of mitochondrial metabolism modulatory properties of ACK1/ATP5F1a signaling and examine ability of (R)-9b to overcome Enzalutamide and abiraterone-resistant CRPCs. The specific aims are as follows: Specific Aim 1. Examine the mechanism by which ATP5F1a-phosphorylation regulates its activity in prostate cancer Specific Aim 2. Explore the role of ACK1/ATP5F1a signaling in prostate cancer models Specific Aim 3. Detail in vivo characterization of ACK1/ATP5F1a signaling in enzalutamide and abiraterone- resistance in mouse models of prostate cancer and patient derived xenografts (PDXs)