Mechanisms of secretion and uptake of small extracellular vesicles (sEVs) in non-small cell lung cancer (NSCLC)

项目摘要 小细胞外囊泡（sEV），通常被称为外来体，是短距离和长距离的介导剂。 细胞间通讯癌症分泌sEV有助于转移、化疗耐药性和耐药。 癌症的进展。我们实验室最近未发表的工作揭示了癌症来源的sEV对肿瘤细胞的影响。 促进侵袭和破坏非致瘤上皮细胞的上皮屏障。因此不 令人惊讶的是，癌症中sEV的研究已经引起了人们的关注。事实上，sEV分泌在哺乳动物中上调， 然而，涉及sEV分泌和生物发生失调的特定机制， 癌细胞还未被充分研究。因此，本提案的目标是确定新的机制， 癌源性sEV的分泌和摄取。为了进一步揭示癌症sEV的机制， 动态，该提案将涉及以下创新目标。在目标1中，我们将确定 突变的KRAS，癌症中最常改变的癌基因之一，对sEV的分泌和生物合成的影响。 这一目的将评估突变KRAS通过损害细胞增殖促进sEV分泌的假设。 sEVs生物合成途径。在aim 2中，我们将发现非小细胞肺使用的新的摄取途径， 癌症（NSCLC）来源的sEV进入非致瘤细胞。基于CRISPR的敲除筛选将在 进行鉴定基因，当丢失时，损害NSCLC sEV摄取。通过这些研究，我们旨在 确定外泌体介导的细胞间通讯赋予致癌特性的机制 到宿主细胞。除了在癌症和sEV生物学中确定新的分子机制外，该项目还将在 还包括一份详细的职业发展计划，将为索托提供额外的指导和培训 这将支持她转变为癌症生物学领域的独立研究者。

PROJECT SUMMARY Small extracellular vesicles(sEVs), often referred as exosomes, are short- and long-distance mediators of intercellular communication. Secretion of sEVs by cancer contributes to metastasis, chemoresistance, and progression of cancer. Recent unpublished work in our lab revealed the impact of cancer-derived sEVs in promoting invasion and disrupting the epithelial barrier of non-tumorigenic epithelial cells. Therefore, it is not surprising that the study of sEVs in cancer has gained attention. Indeed, sEV secretion is upregulated in cancer cells; however, specific mechanisms involved in the dysregulation of sEV secretion and biogenesis in cancer cells is understudied. Therefore, the goal of this proposal is to identify new mechanism that modulate secretion and uptake of cancer-derived sEVs. To further uncover the mechanism involved in cancer sEVs dynamics, the proposal will address the following innovative aims. In aim 1, we will determine the impact of mutant KRAS, one the most frequently altered oncogene in cancer, on the secretion and biogenesis of sEVs. This aim will evaluate the hypothesis that mutant KRAS promotes the secretion of sEVs through impairment of the sEVs biogenesis pathway. In aim 2, we will discover novel uptake pathways used by non-small cell lung cancer (NSCLC) derived sEVs to enter non-tumorigenic cells. A CRISPR-based knockout screen will be conducted to identify genes, that when lost, impair NSCLC sEVs uptake. With these studies we aim to determine the mechanisms by which exosome-mediated cell-cell communication confers oncogenic properties to the host cell. In addition to identifying new molecular mechanisms in cancer and sEVs biology this project also includes a detailed career development plan that will provide Ms. Soto additional mentoring and training that will support her transition into an independent investigator in the cancer biology field.