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（由适用提供）：我的长期职业目标是成为一名独立的学术研究人员，在肌肉生理学领域做出了重大贡献，这些贡献被转化为患有肌肉疾病患者的临床相关疗法。这种奖励机制将有助于通过提供过渡到独立所必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研究了肌肉纤维化如何导致功能和再生受损的杰出问题。为了充分解决这个问题，我已经获得了对细胞外基质特性反应的生物工程的支持，Discher博士。我的同事Barton博士将补充他的精神训练，他将继续在肌肉生理学方面提供出色的精神训练。 Soslowsky博士将提供进一步的指导，Soslowsky博士将在我的咨询委员会任职。该委员会将定期开会，以确保在项目里程碑方面取得足够的进步，在必要时评估替代策略，并监视我的过渡到独立的过渡。我还将与合作者合作； Iwamoto博士在严重的肌肉损伤模型中拥有专业知识和Yamauchi博士，后者是胶原蛋白转录后修改的权威。宾夕法尼亚州的环境非常适合进行拟议的研究，这在很大程度上是宾夕法尼亚肌肉研究所，该研究所提供了对各种肌肉各个方面的敬业教师的各种培训机会。在奖励的修订阶段的教学法将扩大我的研究范围，并在我的职业生涯中尝试新方法的障碍。宾夕法尼亚州还为博士后获得教师职位的支持服务，我将通过生物医学博士后计划办公室充分利用这些职位。宾夕法尼亚大学的杰出机构环境将通过佛罗里达大学与佛罗里达大学Barton博士和北卡罗来纳大学的合作者Yamauchi博士进行培训。提出的这项研究的结果将通过同行审查的出版物和与相关利益的国家会议上的同行审查的出版物和演讲与公众共享。虽然我会领导 项目，订婚的支持系统将确保成功获得我的目标的一切机会。纤维化是组织内细胞外基质（ECM）成分的病理积累，导致结构中断和功能丧失。骨骼肌会响应许多疾病，包括肌肉营养不良和严重的肌肉损伤。在骨骼肌内，ECM不仅提供了细胞支架，而且具有肌肉纤维产生的传输力的额外作用，使ECM对肌肉功能至关重要。我过去在骨骼肌方面的研究表明，在许多情况下，纤维化如何损害肌肉，但也强调了对纤维化材料结构知之甚少。 纤维化组织主要由原纤维胶原蛋白组成，但是胶原蛋白的量不能确定损伤程度，这表明胶原蛋白组织的进一步参数起着重要作用。由于目前尚无批准治疗骨骼肌纤维化的治疗方法，因此我的长期目标是创建治疗靶标和用于测试骨骼肌中抗教纤维的框架工作。我的中心假设是，胶原蛋白组织的参数因纤维化而破坏，包括交联，对准和填料，并且这种破坏会损害肌肉功能和再生。我还将检验以下假设：抑制纤维化肌肉的交联将减少纤维化，改善功能和再生。我的目标是定义骨骼肌纤维化的胶原蛋白组织的新参数，并操纵该组织以探测肌肉再生并提供治疗靶标。在赠款的K99部分期间，我将追求目标1：确定胶原蛋白组织如何改变肌肉纤维化。我们将确定ECM组织的参数，胶原蛋白交联，胶原蛋白对齐和胶原蛋白包装与主动和被动肌肉功能有关。我们将检验以下假设：胶原蛋白组织不仅在骨骼肌纤维化中改变了胶原蛋白。在奖励的修订阶段，我将获得矩阵工程和干细胞生物学方面的新技能，以便在R00阶段AIM 2中购买：确定胶原蛋白组织如何影响卫星细胞再生。我假设对胶原蛋白组织中断的底物上的卫星细胞成熟将受到损害。最后，在K99期间，我将优化一种在R00期间进行测试的治疗方案，从而导致目标3：确定抑制胶原蛋白交联是否导致功能改善和肌肉纤维化的再生。我将检验以下假设：β-氨基原二硫酸酯是一种交联抑制剂，它可以作为抗纤维化治疗有效。这些研究将利用我以前在肌肉生理学方面的研究专业知识，但也需要培训肌肉胶原蛋白和组织培养中基质底物的工程。在该项目的结论中，我将能够定义胶原蛋白组织在骨骼纤维化中的关键特征 肌肉及其与肌肉功能和卫星细胞再生的关系。我还将提供靶向胶原蛋白组织作为治疗骨骼肌纤维化的潜在疗法的证据。这项研究将为我作为独立科学家的职业奠定基础，由该奖项提供的指导培训使我成为可能。