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Roles for increased intracellular pH and heterogeneity in ca

细胞内 pH 值增加和异质性的作用

基本信息

批准号：
10002425
负责人：
Katharine Alice White
金额：
$ 234.75万
依托单位：
UNIVERSITY OF NOTRE DAME
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-05-31
项目状态：
未结题

项目摘要

Abstract One of the greatest challenges in cancer biology is how to overcome issues of tumor heterogeneity and a dynamic microenvironment to select effective therapeutic treatments, reduce therapy resistance, and produce better patient outcomes. While genetic sequencing as standard of care for solid tumors has expanded the therapeutic toolbox, the molecular mechanisms underlying tumor heterogeneity, metastasis, and drug resistance remain poorly understood. For example, failures of targeted therapies like PI3 kinase inhibitors highlight the need for better understanding of how genetic and phenotypic heterogeneity work in concert with microenvironment pressures to enable cancer cell survival, metastasis, and evolution. Although less well studied in terms of heterogeneity, increased intracellular pH (pHi) is a feature of most cancers and enables a host of cancer phenotypes, including increased cell proliferation, metastasis, evasion from apoptosis, migration, and drug resistance. While the constitutively higher pHi of cancer has been shown to enable these behaviors on a population level in various models, little is known about how single-cell spatiotemporal pHi dynamics or pH heterogeneity might influence or drive single-cell cancer phenotypes. Here, I will use an innovative optogenetic tool to spatiotemporally manipulate pHi in living cells and elucidate the role of pHi dynamics in initiating or supporting single-cell cancer behaviors. I hypothesize that increased pHi is a critical indicator of cancer cell function and directly relevant to promoting single-cell invasion and drug resistance. Furthermore, I predict that pHi heterogeneity correlates with other more cryptic markers of heterogeneity including metabolic changes, stem cell markers, and epithelial and mesenchymal markers. In this work, I propose the rigorous investigation of the roles of pHi in supporting cancer cell behaviors through two complementary approaches. First, I will use an optogenetic tool to increase pHi in single cells to determine whether increased pHi is sufficient to drive single-cell invasion and drug resistance in 2D and 3D cancer models. Second, I will obtain pHi heterogeneity maps of 3D cancer models to determine if monitoring pHi increases can be predictive of individual cells that are likely to migrate, invade, or acquire drug resistance. The ability to dynamically measure pHi in living cells makes pHi an attractive biomarker for aggressive tumor subpopulations. Completion of these studies will transform our understanding of how pH dynamics support and promote cancer progression while revealing new routes for monitoring pHi as a diagnostic or prognostic tool or for identifying appropriate therapeutic interventions.

摘要：癌症和生物学研究面临的最大挑战之一是如何克服肿瘤和异质性的问题，以及如何解决这些问题。需要动态的微环境来选择有效的治疗方法，减少药物治疗的耐药性，减少细菌的产生。更好的患者预后。尽管基因测序已成为实体肿瘤患者护理的标准方案，但它的规模也在不断扩大。治疗药物工具箱，包括潜在的分子生物学机制：肿瘤的异质性、肿瘤转移、药物治疗和药物治疗。对耐药性仍然知之甚少。例如，许多靶向治疗的失败，如PI3蛋白激酶抑制剂。强调需要更好地理解遗传基因和表型基因异质性是如何在演唱会上发挥作用的。微环境和压力使癌细胞能够在生存、转移、转移和进化过程中发挥作用。从细胞异质性的角度研究，细胞内pH值(Phi)升高是大多数癌症的一个特征，并使其成为可能。宿主的癌症包括表型，包括细胞增殖增加，转移，逃避细胞凋亡，。迁移、转移和耐药。虽然癌症的PHI值在体质上较高，但尚未有证据表明，这可能会使这些疾病发生。在各种生物模型中，行为依赖于不同的种群水平，但对于单细胞如何影响时空分布Phi，人们知之甚少。动力学或pH值的异质性可能不会影响或驱动单细胞癌的表型。在这里，我将继续使用它。创新的光遗传学工具可以在时空上操纵活细胞中的Phi基因，并阐明Phi基因的主要作用。动力学在启动或支持单细胞癌症的行为方面起到了作用。我假设，增加的PHI是一个关键因素。肿瘤细胞功能的指标直接关系到促进肿瘤单细胞侵袭和耐药的相关指标。此外，我还可以预测，Phi的异质性与其他更神秘的异质性的标记之间存在相关性。包括代谢性细胞变化、干细胞标志物、上皮样细胞和间质干细胞标志物。建议通过这两项调查，更严格地调查他在支持癌症和细胞免疫行为方面所发挥的作用。两种方法相辅相成。首先，我将继续使用一种先进的光遗传学工具来进一步提高单个核细胞的phi水平，以进一步确定。 PHI的增加是否足以推动单细胞肿瘤的侵袭，以及在二维和三维癌症中对药物的耐药性。模型。其次，我将无法获得所有3D癌症模型的Phi异质性和地图，以确定是否需要监测Phi。这些增加也可以作为个体干细胞的预测性指标，因为这些细胞很可能发生迁移、入侵、破坏或获得耐药性。在活体细胞中动态测量Phi的能力使Phi成为一种非常有吸引力的肿瘤侵袭性生物标记物。这些研究的完成将改变我们对pH值和动力学如何支持环境和环境的理解。促进癌症的进展，同时还揭示了用于进一步监测Phi的新的治疗路线，作为一种新的诊断工具或预后评估工具。用于确定适当的治疗性医疗干预措施。