Collagen Proteostasis in Heath and Disease

健康和疾病中的胶原蛋白稳态

• 批准号: 10928439
• 负责人: Matthew Donald Shoulders
• 金额: $ 22.56万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10928439
• 关键词: Address; Affect; Animals; Attention; Basement membrane; Biochemical; Biochemistry; Biological Models; Cartilage; Cells; Chemicals; Chronic stress; Clinical; Collagen; Collagen Diseases; Cystic Fibrosis; Data; Defect; Deposition; Disease; Endoplasmic Reticulum; Extracellular Matrix; Face; Failure; Fibrillar Collagen; Foundations; Functional disorder; Genes; Genetic; Genetic Diseases; Health; Human; Human body; In Vitro; Intervention; Knowledge; Lead; Lectin; Life; Missense Mutation; Modality; Modeling; Modification; Molecular; Molecular Chaperones; Mutation; Nature; Other Genetics; Palliative Care; Pathologic; Pathway interactions; Phenotype; Physiological; Play; Polysaccharides; Preclinical Testing; Process; Procollagen; Production; Property; Proteins; Quality Control; Rapid screening; Research; Skin; Structure; Symptoms; System; Tendon structure; Testing; Therapeutic; Tissue Model; Tissues; Work; autosome; bone; effective therapy; expectation; fundamental research; in vivo; malformation; novel therapeutic intervention; pharmacologic; pre-clinical; protein aggregation; protein misfolding; proteostasis; regenerative therapy; scaffold; screening; small molecule; targeted treatment; triple helix

Collagen is the most abundant protein in animals, constituting up to one-third of total protein in humans. As the major proteinaceous component of tissues ranging from bone and skin to cartilage and basement mem- branes, it constitutes the molecular scaffold for animal life. This ubiquitous protein is challenging for cells to produce, requiring highly coordinated intracellular processes of folding, assembly, and quality control. Owing to the hierarchical nature of collagenous extracellular matrices, the physical and biochemical properties of these tissues are fundamentally defined by these upstream, intracellular processes. Defects, whether genetic or oth- erwise, that are detrimental to any aspect of collagen proteostasis can impact cell health or function and propa- gate to extracellular matrices, leading to diseases known as the collagenopathies. Unfortunately, these dis- eases almost universally lack effective, disease-modifying therapies. Current therapeutic strategies for the collagenopathies focus on regenerative therapies, efforts to strengthen the extracellular matrix itself, or palliative care. None of these approaches aim to address the up- stream issue leading to disease: a failure to properly fold and quality control collagen molecules themselves. If the breakdown of collagen proteostasis could be effectively addressed, the downstream symptoms targeted by current clinical strategies would be alleviated. Indeed, proteostasis enhancement has proven remarkably effi- cacious in many other genetic disorders, including cystic fibrosis, but it has yet to make serious inroads in the collagenopathies. One obstacle is inadequate understanding of the critical decision points in the collagen pro- teostasis network. Another issue is the challenge of pre-clinical testing of proteostasis-targeted interventions in a disease that requires robust, yet biochemically amenable, tissue model systems for discovery efforts. This R01 proposal seeks to address these knowledge gaps, both identifying and elucidating key mech- anisms of intracellular folding and quality control and assessing the therapeutic potential of proteostasis net- work-targeted therapeutics in the collagenopathies. In Aim 1, the functions of the highly conserved procollagen N-glycan in promoting folding, enabling quality control, and identifying when folded procollagens are ready for secretion will be elucidated, via comprehensive work both in cells and in vivo that will reveal the molecular mechanisms of collagen glycoproteostasis. The expectation is demonstration that the long-ignored procollagen N-glycan is actually the critical fulcrum of collagen proteostasis. In Aim 2, a state-of-the-art cartilage-in-a-dish model system is deployed to enable robust testing of proteostasis network-targeted therapies for the colla- genopathies. Combined with mechanistic studies to elucidate the biochemistry of dysregulated collagen prote- ostasis, work in this Aim will provide a strong foundation for a new, proteostasis-focused perspective on treat- ing the collagenopathies.

胶原蛋白是动物中最丰富的蛋白质，占人体总蛋白质的三分之一。 作为从骨骼和皮肤到软骨和基底膜的组织的主要蛋白质成分， 膜，它构成了动物生命的分子支架。这种无处不在的蛋白质对细胞来说是一个挑战， 生产，需要高度协调的折叠，组装和质量控制的细胞内过程。由于 胶原性细胞外基质的分级性质，这些细胞外基质的物理和生化性质， 组织基本上由这些上游的细胞内过程限定。缺陷，无论是遗传性的还是其他的 反之，对胶原蛋白稳定的任何方面都有害的蛋白质会影响细胞的健康或功能， 细胞外基质的大门，导致被称为胶原病的疾病。不幸的是，这些不- 疾病几乎普遍缺乏有效的疾病改善疗法。 目前胶原病的治疗策略集中在再生疗法，努力 加强细胞外基质本身，或姑息治疗。这些方法都不是为了解决问题- 流问题导致疾病：未能正确折叠和质量控制胶原蛋白分子本身。如果 可以有效地解决胶原蛋白质稳态的破坏， 目前的临床策略将得到缓解。事实上，蛋白酶抑制增强已被证明是非常有效的， 它在许多其他遗传性疾病，包括囊性纤维化中也很有效，但它还没有在遗传性疾病中取得重大进展。 胶原蛋白病一个障碍是对胶原蛋白原中的关键决策点的理解不足， 稳态网络另一个问题是临床前测试蛋白质沉积靶向干预的挑战， 这种疾病需要强大的，但生物化学上顺从的组织模型系统来进行发现工作。 R 01提案旨在解决这些知识差距，确定并阐明关键机制， 细胞内折叠和质量控制的分析，并评估蛋白质抑制网的治疗潜力， 胶原蛋白病的靶向治疗。在目的1中，高度保守的前胶原蛋白的功能 N-聚糖促进折叠，实现质量控制，并确定折叠的前胶原何时准备好用于 通过在细胞和体内的全面工作，将阐明分泌的分子机制， 胶原蛋白糖蛋白稳态的机制。这一期望证明了长期被忽视的前胶原 N-聚糖实际上是胶原蛋白稳定的关键支点。在Aim 2中，最先进的软骨培养皿 模型系统的部署，使强大的测试蛋白质稳态网络靶向治疗的胶原蛋白， 遗传病结合机制研究，阐明胶原蛋白表达失调的生物化学， ostasis，在这一目标的工作将提供一个新的，以蛋白质为重点的治疗观点， 胶原蛋白病