Mutating E-cadherin in rats to model lobular breast cancer

突变大鼠 E-钙粘蛋白以模拟小叶乳腺癌

• 批准号: 10830164
• 负责人: Yi Li
• 金额: $ 17.46万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-14 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10830164
• 关键词: Award; Breast Cancer Model; CDH1 gene; Cancer Etiology; Carcinogens; Cell Culture Techniques; Cell Membrane Proteins; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Credentialing; DNA Sequence Alteration; Duct (organ) structure; E-Cadherin; Endocrine; Engineering; Estrogen receptor negative; Estrogen receptor positive; Gene Expression; Genes; Genetic; Goals; Histopathology; Human; Injections; Laboratories; Lead; Lobular; Mammary Neoplasms; Mammary gland; Methods; Modeling; Mus; Mutate; Mutation; NF1 gene; Oncology; Organoids; PIK3CA gene; Parents; Proteins; Proto-Oncogenes; Publishing; Rattus; Reporting; Specimen; Testing; Virus; breast tumorigenesis; cancer initiation; clinically relevant; dimethylbenzanthracene; genetic manipulation; genome editing; in vivo; interest; malignant breast neoplasm; mammary; mammary gland development; mouse model; response; treatment strategy; tumor

This application is being submitted in response to the Notice of Special Interest (NOSI) identified as NOT-CA-23-045.”The parent award R01CA271498 is focused on studying rat models of ductal breast cancer, but it does not study invasive lobular breast cancer (ILC), which accounts for up to 40,000 cases yearly in the US. ILC is predominantly estrogen receptor-positive (ER+). Loss of E-cadherin (encoded by the CDH1 gene) is the pathognomonic feature of ILC being lost at the protein level in over 95% of cases, while mutations of CDH1 occur in 65% of cases. Genetic loss of CDH1 has not been engineered into rat mammary gland cells for studying ILC, but we now have the technical capabilities to engineer CDH1 mutations into rat mammary glands. We predict that CDH1 loss will predispose rat mammary glands for ER+ ILC. In this supplement award, we will team up with the Oesterreich-Lee laboratory, which has extensive expertise in studying ILC, using human specimens, cell culture, and organoids of carcinogen-induced rat tumors. The goal is to develop a clinically relevant rat model of ER+ ILC. Two specific aims are pursued: Aim 1: Examine the effect of somatic CDH1 deletion on rat mammary gland development and function. This somatic CDH1 KO will be achieved using our recently published method using intraductal injection of virus for high efficiency in vivo CRISPR- genome editing. Aim 2: Test whether the combination of somatic CDH1 deletion and PIK3CA activation or DMBA treatment results in rat ER+ mammary tumors with a histopathology and gene expression similar to human ILC. We select the combination of CDH1 KO and PIK3CA activation because CDH1 KO alone is unlikely to sufficient to cause cancer and because PIK3CA is the protooncogene most commonly mutated in human breast cancer including ILC.

该申请是为了响应特殊利益通知（NOSI）而提交 非CA-23-045。“父母奖R01CA271498专注于研究导管乳腺癌的大鼠模型， 但它不研究侵入性小叶乳腺癌（ILC），该乳腺癌每年最多占40,000例 我们。 ILC主要是雌激素受体阳性（ER+）。 E-钙粘蛋白的丧失（由CDH1基因编码） 是超过95％的病例在蛋白质水平上丢失的ILC的病理特征，而突变的突变 CDH1发生在65％的病例中。 CDH1的遗传丧失尚未设计到大鼠乳腺细胞中 研究ILC，但我们现在具有将CDH1突变设计到大鼠乳腺的技术能力 腺体。我们预测CDH1损失将使ER+ ILC诱发大鼠乳腺。在此补充奖中， 我们将与Oesterreich-Lee实验室合作，该实验室在研究ILC方面具有广泛的专业知识。 致癌物诱导的大鼠肿瘤的人类标本，细胞培养和类器官。目标是开发 ER+ ILC的临床相关大鼠模型。追求两个具体目标：目标1：检查躯体的效果 大鼠乳腺发育和功能的CDH1缺失。将实现这个躯体CDH1 KO 使用我们最近发表的方法，使用导管内注射病毒以高效率的体内crispr- 基因组编辑。目标2：测试体细胞CDH1缺失和PIK3CA激活或 DMBA治疗导致具有组织病理学和基因表达的大鼠ER+乳腺肿瘤类似 人ILC。我们选择CDH1 KO和PIK3CA激活的组合，因为仅CDH1 KO是 不可能引起癌症，因为PIK3CA是原子能中最常见的突变 包括ILC在内的人类乳腺癌。