The Role of the Y Chromosome in Bladder Tumor Development, Growth And Progression

Y 染色体在膀胱肿瘤发生、生长和进展中的作用

基本信息

批准号：
10629079
负责人：
DAN THEODORESCU
金额：
$ 54.31万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10629079
关键词：
Address Aftercare Aggressive behavior Alanine Aminopeptidase Animal Model Antibodies Automobile Driving B-Lymphocytes Biological Bladder Bladder Neoplasm CD8-Positive T-Lymphocytes Cancer Patient Catalogs Cell Line Cell model Cells Cessation of life Chromatin Chromosomes Clinical Clustered Regularly Interspaced Short Palindromic Repeats Complement Data Data Set Development Diagnosis Diagnostic Disease Exposure to Functional disorder Gene Deletion Gene Expression Profile Gene Expression Profiling Gene Structure Genes Growth Immune Immune Evasion Immune response Immune system Immunity Immunocompetent Immunophenotyping Incidence Knockout Mice Knowledge Libraries Macrophage Malignant Neoplasms Malignant neoplasm of urinary bladder Mediating Molecular Mus Mutagenesis Myelogenous Myeloid Cells Natural Killer Cells Neoplasm Metastasis Nitrosamines Normal Cell Operative Surgical Procedures Outcome Patient-Focused Outcomes Patients Phenotype Precision therapeutics Process Prognosis Proteins Proteomics Risk Role Smoking T-Lymphocyte Technology Testing The Cancer Genome Atlas Therapeutic Therapeutic Intervention Tobacco Translating Tumor-associated macrophages Urothelial Cell Urothelium Wild Type Mouse Woman X Chromosome Y Chromosome anti-PD-1 anti-PD1 therapy autosome cancer cell cancer risk cancer type chemical carcinogen chromosome Y loss combinatorial differential expression druggable target effective intervention exhaust functional genomics genetic signature histone demethylase immune cell infiltrate immune checkpoint blockade in vivo inhibitor intravesical loss of function male men mortality risk mosaic analysis mutant neoplastic cell next generation sequencing novel overexpression paralogous gene patient prognosis patient stratification pharmacologic prognostic prognostic indicator programmed cell death protein 1 recombinase sex transcriptome sequencing treatment response tumor tumor growth tumor progression tumor-immune system interactions tumorigenesis

项目摘要

PROJECT 2 – SUMMARY Loss of the Y chromosome (LOY) is associated with increased risk of cancer development and death in men with several cancer types. In bladder cancer (BC), a tobacco related malignancy, LOY is seen in ~30% of tumors, yet the biological relevance of this is unknown. Here we address this Knowledge Gap with the Objective to define the molecular mechanisms responsible for LOY driven growth and progression and have Impact by translating these into effective interventions for bladder and other cancer types. Preliminary Data: We examined TCGA BC RNAseq dataset using an LOY gene signature we developed, and found low signature scores associated with reduced patient survival following surgery. To determine if LOY drives BC growth, we developed lineage related male murine cell lines with (Y+) or w/o (Y-) Y chromosome. Y- tumors grew faster than Y+ tumors in immunocompetent mice and cytometric profiling found the former to have increased immunosuppressive tumor-associated macrophages, CD8+ T cells with exhausted phenotype while latter had increased NK cells. In mice (Rag2-/-;Il2rg-/-) that are devoid of T, B and NK cells, Y+ and Y- tumors grew similarly. Transcriptional profiling found four Y chromosome genes differentially expressed in these tumor types, with chromatin organization genes Uty and Kdm5d being the only ones whose expression stratified outcome in TCGA BC patients. Thus, we genetically deleted these genes in Y+ cells and found enhanced tumor growth only in immunocompetent mice while their overexpression in Y- cells reduced their growth. Hence, we test the Hypothesis that Uty and Kdm5d loss enhance BC growth and progression via creation of an immunosuppressive tumor microenvironment in three Specific Aims. In Aim 1 we determine role of Uty and Kdm5d in tumor development and progression using full and urothelial specific conditional Kdm5d or Uty null mice be exposed to the chemical carcinogen BBN. With Project 3 we test the hypothesis that Uty and its X chromosome paralog Utx (aka, Kdm6a) have independent suppressive effects on BC formation and progression by crossing full and urothelial specific conditional Uty and Utx null mice and exposing them to BBN. In Aim 2 we define how Kdm5d and Uty suppress BC and the role of the immune system in this process using defined LOF functional mutants to define the regions on these proteins responsible for tumor growth suppression. With Project 1, we use spatial proteomics to examine immunological infiltrates in relation to mutant and WT Kdm5d and Uty cancer cell expression and various knockout mice or antibody depletion strategy to define which cellular components of the immune system drive tumor growth differences. In Aim 3 we examine ANPEP, the most upregulated gene in Y- tumors that is also a poor prognostic in BC patients and druggable target, as a candidate autosomal effector gene driving Y- tumor growth. We found Y- tumors are more responsive than Y+ tumors to anti-PD-1 and ANPEP depletion in Y- cells made tumors more sensitive to anti-PD-1. Here we will examine the impact of ANPEP deletion or pharmacologic inhibition on the efficacy of anti-PD-1 immune checkpoint blockade therapy.

项目2 - 摘要 Y染色体（LOY）的丧失与男性癌症发展和死亡的风险增加有关有几种癌症类型。在Blader Cancer（BC）中，与烟草相关的恶性肿瘤，在约30％的肿瘤中看到Loy 然而，这是未知的生物学相关性。在这里，我们解决了这个知识差距，目的是定义负责Loy驱动增长和进展的分子机制，并受到影响将其转化为膀胱和其他癌症类型的有效干预措施。初步数据：我们检查了 TCGA BC RNASEQ数据集使用我们开发的Loy基因签名，并发现低签名分数与手术后患者生存减少有关。为了确定Loy是否驱动BC增长，我们开发了与谱系相关的男性鼠细胞系，带有（Y+）或w/o（y-）y染色体。 Y-肿瘤的生长速度快于Y+肿瘤在免疫能力的小鼠和细胞仪分析中，前者的免疫抑制增加了与肿瘤相关的巨噬细胞，CD8+ T细胞具有耗尽的表型，而晚期则增加了NK细胞。没有T，B和NK细胞的小鼠（rag2 - / - ; il2rg - / - ），y+和y肿瘤的生长相似。转录分析发现在这些肿瘤类型中以染色质组织基因表达的四个Y染色体基因不同 UTY和KDM5D是唯一在TCGA BC患者中表达结果分层结果的人。那，我们在Y+细胞中遗传上删除了这些基因，并发现仅在免疫能力小鼠中增强了肿瘤的生长而它们在Y细胞中的过表达减少了其生长。因此，我们检验了UTY和KDM5D的假设损失通过创建免疫抑制性肿瘤微环境来增强BC的生长和发展三个具体目标。在AIM 1中，我们确定UTY和KDM5D在肿瘤发育和进展中的作用使用尿路和尿路特异性有条件的KDM5D或UTY NULL小鼠暴露于化学致癌BBN。通过项目3，我们检验了uty及其X染色体旁系同型UTX（aka，kdm6a）的假设通过完全和尿路上的特异性跨越对卑诗省的形成和进展的独立抑制作用有条件的UTY和UTX无效小鼠，并将其暴露于BBN。在AIM 2中，我们定义KDM5D和UTY如何抑制 BC和免疫系统在此过程中使用定义的LOF功能突变体定义区域的作用这些蛋白质负责肿瘤生长抑制。使用项目1，我们使用空间蛋白质组学与突变体和WT KDM5D和UTY癌细胞表达以及各种淘汰小鼠或抗体部署策略，以定义免疫系统的哪些细胞成分驱动肿瘤生长差异。在AIM 3中，我们检查了ANPEP，这是Y肿瘤中最新的基因在卑诗省患者和可吸毒靶标的预后也很差，作为候选常染色体效应子基因驱动y- 肿瘤生长。我们发现，Y肿瘤比Y+肿瘤对抗PD-1的反应性更高，而在y-中的ANPEP耗竭细胞使肿瘤对抗PD-1更敏感。在这里，我们将研究ANPEP删除的影响或药理学抑制抗PD-1免疫切除剂封锁疗法的效率。