Cholesterol regulation by the cargo receptor SURF4

货物受体 SURF4 的胆固醇调节

• 批准号: 10633136
• 负责人: Brian T Emmer
• 金额: $ 15.42万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10633136
• 关键词: Adult; Affect; Albumins; Alleles; Amino Acids; Apolipoproteins B; Binding; Biological Assay; Biology; Blood; CRISPR screen; Capsid Proteins; Cardiovascular Agents; Cardiovascular Diseases; Cardiovascular Physiology; Cell secretion; Cells; Cholesterol; Cholesterol Homeostasis; Clustered Regularly Interspaced Short Palindromic Repeats; Co-Immunoprecipitations; Coated vesicle; Code; Complex; Computational Biology; Computer Analysis; Cytoplasmic Vesicles; Data Set; Development; Dissection; Embryo; Endoplasmic Reticulum; Enterobacteria phage P1 Cre recombinase; Epitopes; Flow Cytometry; Fluorescence; Foundations; Future; Generations; Genes; Genetic; Genetic Models; Genetic Research; Goals; Growth; Hepatic; Hepatocyte; Heritability; Human; Human Genome; In Vitro; Individual; Investigation; Knock-out; Libraries; Lipoproteins; Liver; Low Density Lipoprotein Receptor; Low-Density Lipoproteins; Maintenance; Massive Parallel Sequencing; Mediating; Mediator; Membrane; Mentors; Mentorship; Metabolism; Molecular; Mus; Partner in relationship; Pathway interactions; Persons; Phenotype; Physicians; Physiological; Plasma; Play; Postdoctoral Fellow; Prevention; Process; Protein Secretion; Proteins; Public Health; Regulation; Regulatory Pathway; Research; Research Personnel; Risk; Role; Scientist; Series; Signal Transduction; Site; System; Testing; Training; Training Activity; Variant; Viral; Work; cardiovascular disorder risk; career; drug development; experience; experimental study; extracellular; functional genomics; genetic variant; genome editing; genome-wide; hypercholesterolemia; improved; in vivo; insight; lifetime risk; loss of function; mouse genetics; mutant; novel; programs; promoter; protein transport; receptor; recruit; screening; success; therapeutic target; tool; transmission process; uptake

PROJECT SUMMARY/ABSTRACT The regulation of human cholesterol plays a critical role in determining a person’s lifetime risk of cardiovascular disease. Identification of the genes and proteins that underlie this process has improved our fundamental understanding of how cardiovascular disease develops, and led to the development of cardiovascular drugs that have greatly benefited public health. While many key regulators of cholesterol homeostasis have been identified, the majority of the heritable risk of hypercholesterolemia cannot be explained by currently recognized variants, suggesting the presence of additional unknown mediators. Recently, breakthroughs in CRISPR-mediated genome editing and massively parallel sequencing have allowed high-throughput, unbiased functional testing of the entire human genome. During my postdoctoral research, I developed a genome-scale CRISPR screen for novel regulators of the extracellular secretion of PCSK9, a process that plays a critical role in modulating plasma cholesterol levels. This led to my discovery of SURF4 as a cargo receptor that promotes efficient PCSK9 secretion in vitro. However, the physiologic relevance and mechanistic basis of the interaction between SURF4 and PCSK9 remains unclear. This proposal builds upon my prior work through the generation of mice with liver-specific deletion of Surf4 and characterization of the consequences to cholesterol homeostasis (Aim 1), and by altering the coding sequence of human SURF4 to determine which domains mediate its interaction with PCSK9 (Aim 2). In addition to elucidating the basic biology of PCSK9 secretion, these studies will inform the potential of SURF4 as a therapeutic target and serve as proof-of-principle for the power of CRISPR screening to identify novel regulators of protein secretion. This research will be conducted under the guidance of a primary mentor who has extensive expertise in protein trafficking, cardiovascular physiology, and genetics as well as a long track record of mentorship for early career physician-scientists who have successfully transitioned to scientific independence. This work, along with the mentorship and training activities described in my proposal, will facilitate my development toward my ultimate goal of becoming an academic physician-scientist and independent investigator.

项目概要/摘要 人类胆固醇的调节在决定一个人终生患心血管疾病的风险方面起着至关重要的作用 识别这一过程背后的基因和蛋白质已经改善了我们的基础。 了解心血管疾病如何发展，并导致心血管药物的开发 这极大地有益于公众健康，而胆固醇稳态的许多关键调节因子已被研究。 已确定，高胆固醇血症的大部分遗传风险无法用目前的方法来解释 已识别的变异，表明存在其他未知介质。 CRISPR 介导的基因组编辑和大规模并行测序实现了高通量、无偏倚 在我的博士后研究期间，我开发了一个基因组规模的功能测试。 CRISPR 筛选 PCSK9 胞外分泌的新型调节因子，这一过程发挥着关键作用 这导致我发现 SURF4 作为一种促进胆固醇水平的货物受体。 然而，相互作用的生理相关性和机制基础。 SURF4 和 PCSK9 之间的关系仍不清楚。该提案建立在我之前一代的工作基础上。 肝脏特异性删除 Surf4 的小鼠及其对胆固醇后果的表征 稳态（目标 1），并通过改变人类 SURF4 的编码序列来确定哪些结构域 介导其与 PCSK9 的相互作用（目标 2） 除了阐明 PCSK9 分泌的基本生物学之外， 这些研究将揭示 SURF4 作为治疗靶点的潜力，并作为原理验证 这项研究将利用 CRISPR 筛选来鉴定新型蛋白质分泌调节剂。 在主要导师的指导下，他在蛋白质运输、心血管等方面拥有丰富的专业知识 生理学和遗传学，以及对早期职业医师科学家的长期指导记录 在指导和培训的帮助下，我们已成功过渡到科学独立。 我的提案中描述的活动将有助于我朝着成为一名 学术医师科学家和独立研究者。

项目成果

ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes.

同基因细胞系内的 ACE2 蛋白表达是异质的，并且与不同的转录组相关。

• 发表时间: 2021
• 影响因子: 4.6
• 作者: Sherman, Emily J;Emmer, Brian T
• 通讯作者: Emmer, Brian T

Type I interferon signaling induces a delayed antiproliferative response in respiratory epithelial cells during SARS-CoV-2 infection.

I 型干扰素信号传导在 SARS-CoV-2 感染期间诱导呼吸道上皮细胞延迟抗增殖反应。

• 发表时间: 2023-12-21
• 作者: Bragazzi Cunha, Juliana;Leix, Kyle;Sherman, Emily J;Mirabelli, Carmen;Frum, Tristan;Zhang, Charles J;Kennedy, Andrew A;Lauring, Adam S;Tai, Andrew W;Sexton, Jonathan Z;Spence, Jason R;Wobus, Christiane E;Emmer, Brian T
• 通讯作者: Emmer, Brian T

Type I interferon signaling induces a delayed antiproliferative response in Calu-3 cells during SARS-CoV-2 infection.

在 SARS-CoV-2 感染期间，I 型干扰素信号传导可诱导 Calu-3 细胞延迟抗增殖反应。

• 发表时间: 2023-03-01
• 作者: Cunha, Juliana Bragazzi;Leix, Kyle;Sherman, Emily J;Mirabelli, Carmen;Kennedy, Andrew A;Lauring, Adam S;Tai, Andrew W;Wobus, Christiane E;Emmer, Brian T
• 通讯作者: Emmer, Brian T