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Cancer Resistant Mice

抗癌小鼠

基本信息

批准号：
10533357
负责人：
BRUCE A BEUTLER
金额：
$ 66.7万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10533357
关键词：
Ablation Acceleration Affect Alleles Animals Antibodies Antigen Presentation Antigen-Presenting Cells Autoantigens Biological Assay Bone Marrow Transplantation CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes CRISPR/Cas technology Cancer Etiology Cancer Model Chimerism Cloning Code Complex Defect Dendritic Cells Detection Embryo Enrollment Epithelium Ethylnitrosourea Failure Frequencies Generations Genes Genetic Genome Genome Stability Genotype Germ-Line Mutation Histocompatibility Histocompatibility Antigens Class I Histocompatibility Antigens Class II Homozygote House mice Human Hyperplasia Immune Immune response Immune system Immunologics Inbred Mouse Inbred Strain Individual Induced Mutation Knockout Mice MHC Class I Genes MHC Class II Genes Malignant Neoplasms Maps Measures Mediating Meiosis Melanoma Cell Minor Missense Mutation Mus Mutagenesis Mutagens Mutant Strains Mice Mutate Mutation Myelogenous Nitrosourea Compounds Nuclear Nutritional Patients Peptides Phenotype Point Mutation Population Productivity Proteins RNA Splicing Reaction Regulatory T-Lymphocyte Resistance Safety Screening for cancer Site Skin graft T cell therapy T-Lymphocyte Testing Therapeutic Thymus Gland Transcript Translating Tumor Volume Vascularization Work anti-PD1 antibodies autosome cancer genome cancer therapy cancer transplantation causal variant central tolerance clinical application functional mimics genetic pedigree humanized mouse interest male melanoma mutant neoantigens neoplastic cell nonsynonymous mutation novel novel strategies prevent refractory cancer repository resistance allele resistance mechanism resistance mutation screening stem subcutaneous synergism targeted cancer therapy therapeutic development translational applications translational study tumor tumor growth

项目摘要

PROJECT SUMMARY Can germline mutations cause strong resistance to otherwise lethal cancers? Certain germline genotypes might be poorly supportive of tumor vascularization, nutritional demands, or resistance to immune attack, yet compatible with host survival. Of particular interest, some mutations might abet the host response to neo- antigens, or even to self-antigens highly expressed in syngeneic tumors. The identification of resistance mutations could provide new approaches and targets for cancer therapy. At least in human populations, resistance mutations would be very difficult to identify. Human germline genetic variability, stem variability among cancer genomes, and the high frequency of humans who never develop cancer throughout their lives would make mapping novel human resistance alleles all but impossible. In mice, finding such mutations is much easier. Syngeneic tumor lines (with relatively stable genomes) exist for many inbred strains of Mus musculus. The inbred mice themselves have a defined germline reference sequence. Each individual is homozygous at nearly all loci, and almost genetically identical to all others. Over the past several years, we took advantage of this situation to identify genes in which mutations confer cancer resistance. Using the random germline mutagen ENU, we created third generation (G3) germline mutant mice (C57BL/6J strain). A total of 23,751 third-generation (G3) mice from 561 pedigrees, bearing a total of 32,039 non-synonymous coding/splicing changes were enrolled into a screen in which each mouse was injected subcutaneously with 2e5 B16F10 melanoma cells, and anti-PD-1 antibody was administered on days 5, 8, and 11. Tumor volume was measured on days 13 and 20. The G1 male founder of each pedigree was sequenced to identify all non-synonymous coding/splicing mutations induced by mutagenesis, and all G3 descendants were genotyped at all induced mutation sites in advance of screening. Automated meiotic mapping allowed quick detection of even subtle phenotypes and assignment to causative mutations. This screen yielded several mutations causing resistance to transplantable cancers. 14.2% saturation of the autosomal genome was achieved in screening (fraction of autosomal genes with severely damaging or destructive alleles tested in the homozygous state three times or more). Therefore, much remains undiscovered. From what we know already, there is a realistic chance of translating genetic discoveries from this screen to human cancer therapy. This proposal aims to extend screening for cancer resistance, and to further advance mechanistic and translational studies of two resistance mutations, each in a gene with a human orthologue, testing synergy between therapeutic approaches built around each protein target, and laying groundwork for clinical applications.

项目概要种系突变能否对致命的癌症产生强大的抵抗力？某些种系基因型可能对肿瘤血管化、营养需求或对免疫攻击的抵抗力的支持较差，但与宿主生存相适应。特别有趣的是，一些突变可能会促进宿主对新生物的反应。抗原，甚至是在同基因肿瘤中高度表达的自身抗原。耐药性的鉴定突变可以为癌症治疗提供新的方法和靶点。至少在人类群体中，抗性突变将很难识别。人类种系遗传变异、茎变异癌症基因组，以及终其一生从未患癌症的人类的高频率使得绘制新的人类抗性等位基因几乎不可能。在小鼠中，发现此类突变要容易得多。许多小家鼠近交系都存在同基因肿瘤系（具有相对稳定的基因组）。近交系小鼠本身具有明确的种系参考序列。每个个体几乎所有基因座都是纯合的，并且在基因上几乎与所有其他人相同。在过去的几年里，我们利用这种情况识别突变赋予癌症抗性的基因。使用随机种系诱变剂 ENU，我们创建第三代（G3）种系突变小鼠（C57BL/6J品系）。第三代（G3）共计23,751辆来自 561 个谱系、总共具有 32,039 个非同义编码/剪接变化的小鼠被纳入研究每只小鼠皮下注射 2e5 B16F10 黑色素瘤细胞和抗 PD-1 的筛选在第 5、8 和 11 天施用抗体。在第 13 和 20 天测量肿瘤体积。G1 雄性对每个谱系的创始人进行测序，以识别由以下因素引起的所有非同义编码/剪接突变：诱变，并且在筛选前对所有诱导突变位点的所有 G3 后代进行基因分型。自动减数分裂图谱可以快速检测甚至细微的表型并分配致病因子突变。该筛选产生了几种突变，导致对可移植癌症的抵抗。 14.2% 饱和度常染色体基因组的筛选是在筛选中实现的（具有严重破坏性或严重性的常染色体基因的分数）破坏性等位基因在纯合状态下测试三次或更多次）。因此，还有很多东西未被发现。据我们所知，将基因发现从这个屏幕转化为现实的机会人类癌症治疗。该提案旨在扩大癌症耐药性筛查范围，并进一步推进对两种耐药突变的机制和转化研究，每种突变都存在于具有人类直系同源物的基因中，测试围绕每个蛋白质靶点建立的治疗方法之间的协同作用，并为临床应用。