Project 1: Evolution of transcriptional programs in lung cancer progression

项目1：肺癌进展中转录程序的进化

• 批准号: 9356947
• 负责人: TYLER E. JACKS
• 金额: $ 42.08万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9356947
• 关键词: Ablation; Address; Affect; Biological Response Modifiers; CRISPR/Cas technology; Cancer Biology; Cancer Etiology; Candidate Disease Gene; Cells; Cessation of life; Collaborations; DNA Sequence Alteration; Data; Development; Diphtheria Toxin; Enzymes; Epigenetic Process; Evolution; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Genomics; Genotype; HMGA2 gene; Heterogeneity; Human; Immune; Immune Evasion; Immune response; Immunologic Markers; Immunosuppression; Individual; Infiltration; Inflammatory; Investigation; KRAS2 gene; Killer Cells; Laboratories; Lung Adenocarcinoma; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Mediator of activation protein; Messenger RNA; Methods; Modeling; Molecular Profiling; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Pathway interactions; Pharmacology; Phenotype; Population; Porcupines; Prevention; Process; Property; Regional Cancer; Series; Signal Pathway; Signal Transduction; Somatic Mutation; TP53 gene; Technology; Testing; Time; United States; Validation; WNT Signaling Pathway; base; cancer cell; cancer genomics; cancer stem cell; cancer type; chemotherapy; cytokine; disorder subtype; gene function; genetic technology; genome editing; immunogenic; improved; innovation; interest; lens; mouse model; mutant; neoplastic cell; new therapeutic target; programs; response; response biomarker; single cell analysis; stem; stem-like cell; targeted treatment; tumor; tumor microenvironment; tumor progression; Ablation; Address; Affect; Biological Response Modifiers; CRISPR/Cas technology; Cancer Biology; Cancer Etiology; Candidate Disease Gene; Cells; Cessation of life; Collaborations; DNA Sequence Alteration; Data; Development; Diphtheria Toxin; Enzymes; Epigenetic Process; Evolution; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Genomics; Genotype; HMGA2 gene; Heterogeneity; Human; Immune; Immune Evasion; Immune response; Immunologic Markers; Immunosuppression; Individual; Infiltration; Inflammatory; Investigation; KRAS2 gene; Killer Cells; Laboratories; Lung Adenocarcinoma; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Mediator of activation protein; Messenger RNA; Methods; Modeling; Molecular Profiling; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Pathway interactions; Pharmacology; Phenotype; Population; Porcupines; Prevention; Process; Property; Regional Cancer; Series; Signal Pathway; Signal Transduction; Somatic Mutation; TP53 gene; Technology; Testing; Time; United States; Validation; WNT Signaling Pathway; base; cancer cell; cancer genomics; cancer stem cell; cancer type; chemotherapy; cytokine; disorder subtype; gene function; genetic technology; genome editing; immunogenic; improved; innovation; interest; lens; mouse model; mutant; neoplastic cell; new therapeutic target; programs; response; response biomarker; single cell analysis; stem; stem-like cell; targeted treatment; tumor; tumor microenvironment; tumor progression

Project Summary – Project 1 Tumor progression is the product genetic mutations that affect gene function and epigenetic alterations that affect patterns of gene expression and cellular states. Together with the effects of the tumor microenvironment, these changes produce the phenotypes that underlie the biology of cancer. From extensive genomic studies performed over the past decade the genetic landscape of cancer progression is beginning to emerge, but much less is know about how tumors evolve epigenetically. Project 1 is focused on the investigation of tumor evolution through the lens of changes in epigenetic states and gene expression programs. It is anticipated that an improved understanding of these aspects of tumor development will yield new strategies to intervene in this process as well as to treat established tumors. Three Specific Aims will be pursued addressing these questions in the context of well-established genetically-engineered mouse models of human lung adenocarcinoma (LUAD) based on mutations of K-ras (“K”) or the combination of K-ras and p53 (“KP”). Of note, K-ras mutations occur in approximately 30% of human LUAD and targeted therapies do not exist for this subtype of the disease. Preliminary data in support of the Project 1 demonstrates that as tumors progress in the KP model they develop subpopulations of cells that have cancer stem-cell like properties and show markers of response to the Wnt signaling pathway and other cells that markers of Wnt-producing cells that express the enzyme Porcupine (Porcn). It is hypothesized that these Porcn+ cells act as niche cells for the cancer stem-like cells. In Aim 1, a combination of genetic marking, molecular profiling and cell ablation studies will be used to characterized and functionally test these putative niche cells for their importance in tumor development. These studies may identify new targets for the treatment of human lung cancer. Using single cell mRNA sequencing (SCmRNAseq), performed in collaboration with the laboratory of Aviv Regev, the Jacks laboratory has profiled individual cancer cells from K and KP tumors. These data reveal extensive intratumoral and intertumoral transcriptional heterogeneity. Aim 2 of Project 1 will complete the characterization of transcriptional heterogeneity as a function of tumor genotype and tumor progression as well as explore the epigenetic mediators of these effects. Finally, the transcriptional programs of cancer cells are expected to be affected by immune infiltration and anti-tumor immune responses, potentially in a heterogeneous fashion across tumors. These questions will be explored in Aim 3 of Project 1 using SCmRNAseq-based profiling and functional studies of “immunogenic and “nonimmunogenic” tumors. The identification and functional validation of pathways that contribute to immune suppression in this model could reveal new targets of therapy for human lung cancer and other cancer types. In all of the Aims of Project 1, a series of innovative genetic technologies (including lineage tracing, cell marking, cell ablation, and CRISPR/Cas9-mediated genome editing and gene activation) will be used to functionally characterization of genes and pathways identified through profiling.

项目摘要 - 项目1 肿瘤进展是影响基因功能和表观遗传改变的产物基因突变 影响基因表达和细胞态的模式。以及肿瘤微环境的影响 这些变化产生了癌症生物学基础的表型。来自广泛的基因组研究 在过去的十年中，癌症进展的遗传景观开始出现，但是 更少知道肿瘤如何表观发展。项目1专注于肿瘤的研究 通过表观遗传状态和基因表达程序的变化镜头的演变。预计 对肿瘤发育的这些方面的改进理解将产生新的策略来干预这一点 过程以及治疗已建立的肿瘤。将追求三个具体目标，以解决这些问题 在人类肺腺癌的良好成熟的一般工程小鼠模型的背景下 （LUAD）基于K-Ras（“ K”）的突变或K-Ras和p53（“ KP”）的组合。值得注意的是，k-ras突变 大约30％的人类LUAD和靶向疗法发生在这种亚型的亚型中 疾病。支持项目1的初步数据表明，随着KP模型中肿瘤的进展 他们开发具有癌症类似于特性的细胞的亚群，并显示了反应的标记 到Wnt信号通路和其他表达酶的产生Wnt的细胞的细胞 豪猪（猪）。假设这些Porcn+细胞充当癌症干细胞的小裂细胞。在 AIM 1，将使用遗传标记，分子分析和细胞消融研究的组合来 表征并在功能上测试了这些假定的小众细胞在肿瘤发育中的重要性。这些 研究可能会确定治疗人肺癌的新目标。使用单细胞mRNA测序 （SCMRNASEQ）与Aviv Regev实验室合作进行，Jacks实验室已经进行了介绍 来自K和KP肿瘤的单个癌细胞。这些数据揭示了广泛的肿瘤内和室间 转录异质性。项目1的目标2将完成转录的表征 异质性与肿瘤基因型和肿瘤进展的关系以及探索表观遗传 这些影响的介体。最后，预计癌细胞的转录程序会受到 免疫浸润和抗肿瘤免疫反应，可能以各种肿瘤的异质方式以异质性。 这些问题将在项目1的AIM 3中使用基于SCMRNASEQ的分析和功能探索 研究“免疫原性和“非免疫原性”肿瘤。 在该模型中有助于免疫抑制的途径可以揭示人类治疗的新靶标 肺癌和其他癌症类型。在项目1的所有目标中，一系列创新的遗传技术 （包括谱系跟踪，细胞标记，细胞消融以及CRISPR/CAS9介导的基因组编辑和基因 激活将用于在功能上表征通过分析识别的基因和途径。