The role of collagen organization in determination of fibrotic muscle function and regeneration

胶原组织在纤维化肌肉功能和再生测定中的作用

• 批准号: 9274036
• 负责人: LUCAS R SMITH
• 金额: $ 9.29万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9274036
• 关键词: Acute; Address; Adverse effects; Advisory Committees; Affect; Alpha Cell; Architecture; Award; Basement membrane; Biochemical; Biology; Biomedical Engineering; Cell Differentiation process; Cell Maturation; Cell Proliferation; Cessation of life; Cicatrix; Collagen; Complement; Deposition; Engineering; Ensure; Environment; Extracellular Matrix; Extracellular Structure; Faculty; Fibrillar Collagen; Fibrosis; Florida; Foundations; Genetic Transcription; Goals; Grant; Impairment; In Vitro; Injury; Institutes; Knowledge; LOX gene; Leadership; Maintenance; Measures; Mentors; Mentorship; Modeling; Modification; Monitor; Muscle; Muscle Cells; Muscle Fibers; Muscle function; Muscular Atrophy; Muscular Dystrophies; Myopathy; Natural regeneration; Nature; North Carolina; Pathologic; Peer Review; Pennsylvania; Phase; Play; Polarization Microscopy; Positioning Attribute; Postdoctoral Fellow; Prevention; Production; Property; Protein-Lysine 6-Oxidase; Publications; Research; Research Personnel; Role; Scientist; Services; Skeletal Muscle; Solubility; Structure; Support System; Techniques; Testing; Therapeutic; Tissues; Training; Training Programs; Translating; Treatment Protocols; Universities; Weight-Bearing state; Work; authority; base; beta Aminopropionitrile; career; cell motility; clinically relevant; crosslink; density; effective therapy; experience; improved; improved functioning; in vivo; inhibitor/antagonist; interest; loss of function; migration; mouse model; muscle physiology; muscle regeneration; novel strategies; novel therapeutics; premature; programs; public health relevance; regenerative; repaired; response; satellite cell; scaffold; skills; stem cell biology; success; symposium; therapeutic target; tissue culture; training opportunity

 DESCRIPTION (provided by applicant): My long term career goal is to become an independent academic researcher, making substantial contributions in the field of muscle physiology that are translated into clinically relevant therapies for those with muscle diseases. This award mechanism would be instrumental in the providing the ability to reach goal by providing the continued training necessary for me to transition to independence. My background in bioengineering and muscle physiology research has provided a focus on outstanding question of how muscle fibrosis leads to impaired function and regeneration. To adequately address this question I have garnered the support of an outstanding mentor in bioengineering of cellular responses to extracellular matrix properties, Dr. Discher. His mentorship will be supplemented by that of my co-mentor Dr. Barton, who will continue to provide exceptional mentorship in muscle physiology. Further guidance will be provided by Dr. Soslowsky, who will serve on my advisory committee. This committee will meet regularly to ensure adequate progress toward project milestones, assess alternative strategies when necessary, and monitor my transition to becoming independent. I will also work with collaborators; Dr. Iwamoto who has expertise in severe muscle injury models and Dr. Yamauchi who is an authority on collagen post-transcriptional modifications. The environment at Penn is ideal for conducting the proposed research, in large part from the Pennsylvania Muscle Institute which provides various training opportunities among a cadre of engaged faculty interested in a variety of aspects of muscle. Didactics during the mentored phase of the award will broaden the scope of my research and lower barriers to attempting new approaches in my career. Penn also provides support services for postdocs acquiring faculty positions that I will take full advantage of through the Biomedical Postdoctoral Program Office. The outstanding institutional environment at the University of Pennsylvania will be supplemented by training at the University of Florida with co-mentor Dr. Barton and at the University of North Carolina with collaborator Dr. Yamauchi. The results of this research proposed will be shared with the public through peer reviewed publications and presentations at national conferences with relevant interests. While I will take leadership of this project, the support system engaged will ensure every opportunity for success in acquiring my goals. Fibrosis is the pathologic accumulation of extracellular matrix (ECM) components within a tissue leading to disrupted architecture and loss of function. Skeletal muscle undergoes fibrosis in response to many conditions including muscular dystrophies and severe muscle injury. Within skeletal muscle the ECM not only provides a cell scaffold, but has the additional role of transmitting forces produced by muscle fibers, making ECM critical to muscle function. My past research in skeletal muscle has demonstrated how fibrosis compromises muscle in many conditions, but also highlighted how little is known about the structure of fibrotic material. Fibrotic tissue is primarily made of up fibrillar collagen, however the amount of collagen does not determine the degree of impairment, suggesting further parameters of collagen organization play an important role. As there is currently no approved therapeutic to treat skeletal muscle fibrosis, my long term goal is to create therapeutic targets and a frame work for testing anti-fibrotics in skeletal muscle. My central hypothesis is that parameters of collagen organization are disrupted in fibrosis, including cross-linking, alignment, and packing, and that this disruptio impairs muscle function and regeneration. I will also test the hypothesis that inhibiting cross-linking in fibrotic muscle will reduce fibrosis, improve function, and regeneration. My objective i to define new parameters of collagen organization of skeletal muscle fibrosis and manipulate that organization in order to probe muscle regeneration and provide therapeutic targets. During the K99 portion of the grant I will pursue Aim 1: Determine how collagen organization is altered in muscle fibrosis. We will determine how parameters of the ECM organization, collagen cross-linking, collagen alignment, and collagen packing relate to active and passive muscle function. We will test the hypothesis that collagen organization, not just collagen quantity, is altered in skeletal muscle fibrosis. During the mentored phase of the award I will obtain new skills in matrix engineering and stem cell biology in order to pursue in the R00 phase Aim 2: Determine how collagen organization effects satellite cell regeneration. I hypothesize satellite cell maturation on substrates with disrupted collagen organization will be impaired. Finally during the K99 I will optimize a treatment regimen to be tested during the R00 phase leading to Aim 3: Determine if inhibiting collagen cross-linking leads to improved function and regeneration of muscle fibrosis. I will test the hypothesis that β-aminopropionitrile, a cross-linking inhibitor, an be effective as an anti-fibrotic treatment. These studies will take advantage of my previous research expertise in muscle physiology, but also require training in the analysis of muscle collagen and the engineering of matrix substrates in tissue culture. At the conclusion of this project, I will be able to define the key features of collagen organization in fibrosis of skeletal muscle and their relation to muscle function and satellite cell regeneration. I will also provide evidence for targeting collagen organization as a potential therapy to treat skeletal muscle fibrosis. This research will lay the foundation for my career as an independent scientist made possible by the mentored training provided by this award.

 描述（由申请人提供）：我的长期职业目标是成为一名独立的学术研究员，在肌肉生理学领域做出重大贡献，并将其转化为针对肌肉疾病患者的临床相关疗法。通过为我提供过渡到独立所需的持续培训来提供实现目标的能力。我在生物工程和肌肉生理学研究方面的背景使我重点关注肌肉纤维化如何导致功能和再生受损的突出问题。我已获得细胞对细胞外基质特性的生物工程领域的杰出导师迪舍尔博士的支持将得到我的合作导师巴顿博士的补充，巴顿博士将继续在肌肉生理学方面提供卓越的指导。由 Soslowsky 博士提供，他将担任我的顾问委员会成员，该委员会将定期举行会议，以确保项目里程碑取得足够进展，在必要时评估替代策略，并监督我向独立的转变。 . Iwamoto 拥有严重肌肉损伤模型方面的专业知识，而 Yamauchi 博士是胶原蛋白转录后修饰方面的权威。宾夕法尼亚大学的环境非常适合进行拟议的研究，其中大部分来自宾夕法尼亚肌肉研究所，该研究所为这些人提供各种培训机会。在该奖项的指导阶段，一批对肌肉各个方面感兴趣的敬业教师将扩大我的研究范围，并降低我在职业生涯中尝试新方法的障碍。宾夕法尼亚大学还为博士后获得教师职位提供支持服务。我会采取通过生物医学博士后项目办公室的充分利用，宾夕法尼亚大学出色的机构环境将通过在佛罗里达大学与合作导师巴顿博士和在北卡罗来纳大学与合作者山内博士的培训得到补充。这项研究的建议将通过同行评审的出版物和在有相关兴趣的全国会议上的演讲与公众分享，而我将领导这项工作。 项目中，参与的支持系统将确保成功实现我的目标的每一个机会。 纤维化是组织内细胞外基质（ECM）成分的病理性积累，导致骨骼肌在许多情况下发生纤维化。在骨骼肌中，ECM 不仅提供细胞支架，还具有传递肌纤维产生的力的额外作用，这使得 ECM 对肌​​肉功能至关重要。骨骼肌已经证明了纤维化如何在许多情况下损害肌肉，但也强调了人们对纤维化材料的结构知之甚少。 纤维化组织主要由纤维状胶原组成，但胶原蛋白的量并不能决定损伤的程度，这表明胶原组织的进一步参数发挥着重要作用，因为目前还没有批准的治疗方法来治疗骨骼肌纤维化，从长远来看。我的目标是创建治疗靶标和测试骨骼肌抗纤维化的框架，我的中心假设是胶原组织的参数在纤维化过程中被破坏，包括交联、排列和堆积，并且这种破坏会破坏骨骼肌的抗纤维化。我还将测试抑制纤维化肌肉中的交联将减少纤维化、改善功能和再生的假设，我的目标是定义骨骼肌纤维化的胶原蛋白组织的新参数并按顺序操纵该组织。探索肌肉再生并提供治疗目标 在拨款的 K99 部分中，我将追求目标 1：确定胶原蛋白组织在肌肉纤维化中如何改变 我们将确定 ECM 组织、胶原蛋白的参数如何变化。交联、胶原蛋白排列和胶原蛋白堆积与主动和被动肌肉功能相关，我们将测试骨骼肌纤维化中胶原蛋白组织（而不仅仅是胶原蛋白数量）发生变化的假设，在获奖阶段我将获得导师。基质工程和干细胞生物学的新技能，以便在 R00 阶段实现目标 2：确定胶原组织如何影响卫星细胞再生优化在 R00 阶段测试的治疗方案，实现目标 3：确定抑制胶原蛋白交联是否会改善肌肉纤维化的功能和再生。这些研究将利用我之前在肌肉生理学方面的研究专业知识，但还需要在肌肉胶原蛋白分析和组织培养中基质基质工程方面进行培训。在这个项目中，我将能够定义骨骼纤维化中胶原组织的关键特征 我还将提供证据，证明以胶原组织为治疗骨骼肌纤维化的潜在疗法，这项研究将为我作为一名独立科学家的职业生涯奠定基础，而这种职业生涯是通过指导培训实现的。由该奖项提供。