Cancer Resistant Mice

抗癌小鼠

基本信息

批准号：
10364495
负责人：
BRUCE A BEUTLER
金额：
$ 68.06万
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/10364495
关键词：
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 Epithelial Ethylnitrosourea Failure Frequencies Generations Genes Genetic Genome 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 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 Proteins RNA Splicing Regulatory T-Lymphocyte Resistance Safety Screening for cancer Site Skin graft T cell therapy T-Lymphocyte Testing Therapeutic Thymus Gland Time Transcript Translating Tumor Volume Vascularization Work anti-PD1 antibodies autosome base cancer genome cancer therapy cancer transplantation causal variant central tolerance clinical application 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

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 Epithelial Ethylnitrosourea Failure Frequencies Generations Genes Genetic Genome 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 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 Proteins RNA Splicing Regulatory T-Lymphocyte Resistance Safety Screening for cancer Site Skin graft T cell therapy T-Lymphocyte Testing Therapeutic Thymus Gland Time Transcript Translating Tumor Volume Vascularization Work anti-PD1 antibodies autosome base cancer genome cancer therapy cancer transplantation causal variant central tolerance clinical application 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

展开

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.

项目摘要种系突变会引起对其他致命癌的强烈抗药性吗？某些种系基因型可能对肿瘤血管形成，营养需求或对免疫攻击的抵抗力不大，但与宿主生存兼容。特别有趣的是，某些突变可能会教宿主对新的反应抗原，甚至是在合成肿瘤中高度表达的自我抗原。抵抗的识别突变可以为癌症治疗提供新的方法和靶标。至少在人口中，阻力突变将非常难以识别。人类种系遗传变异性，茎变异性癌症基因组和一生从未发展癌症的人类的高频率将会使映射新颖的人类抵抗等位基因几乎不可能。在小鼠中，找到这种突变要容易得多。许多近交菌株的杂种菌株存在合成性肿瘤系（具有相对稳定的基因组）。近交小鼠本身具有定义的种系参考序列。每个人几乎在所有基因座上都是纯合的，几乎与其他所有人相同。在过去的几年中，我们利用这种情况确定突变赋予癌症抗性的基因。使用随机种系诱变剂ENU，我们创建的第三代（G3）种系突变小鼠（C57BL/6J菌株）。总共23,751第三代（G3）来自561个血统的小鼠总共招募了32,039个非同义编码/拼接更改将每只小鼠皮下注入2E5 B16F10黑色素瘤细胞和抗PD-1 在第5、8和11天给予抗体。在第13和20天测量肿瘤体积。对每个血统的创始人进行了测序，以识别由由诱变，所有G3后代在筛查之前均在所有诱导的突变位点进行基因分型。自动化的减数分裂映射允许快速检测甚至微妙的表型和分配为因果关系突变。该屏幕产生了几种突变，导致对移植癌症的抗性。 14.2％饱和度在筛查中实现了常染色体基因组的（具有严重破坏或严重破坏的常染色体基因的比例）在纯合状态测试的破坏性等位基因三次或更多）。因此，仍然没有发现。据我们所知，将遗传发现从此屏幕转化为现实的机会人类癌症治疗。该建议旨在扩大筛查癌症的耐药性，并进一步促进两个抗性突变的机械和翻译研究，每个都有人类直系同源物的基因测试围绕每个蛋白质目标建立的治疗方法之间的协同作用，并为临床应用。