Determining the molecular organization of TDP-43 in liquid spherical annuli and establishing a designer gene therapeutic strategy to degrade TDP-43 aggregates

确定液体球形环中 TDP-43 的分子组织并建立降解 TDP-43 聚集体的设计基因治疗策略

• 批准号: 10589577
• 负责人: Haiyang Yu
• 金额: $ 24.9万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-21 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10589577
• 关键词: Affinity; Alternative Splicing; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Antibodies; Antisense Oligonucleotides; Authorship; Award; Binding; Brain; Cell Line; Cell Nucleus; Cell model; Cells; Clinical Trials; Collaborations; Consensus; Cryo-electron tomography; Cryoelectron Microscopy; Cytoplasm; Cytoplasmic Protein; Dependovirus; Electron Microscopy; Environment; Excision; Failure; Foundations; Genes; Goals; Human; Immunoglobulin Fragments; In Situ; In Vitro; Lead; Letters; Liquid substance; Mediating; Mentors; Methods; Minor; Modeling; Molecular; Motor; Motor Neurons; Mus; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Nuclear; Nuclear RNA; Pathologic; Pathology; Pathway interactions; Phase; Physiological; Process; Productivity; Property; Proteins; Publications; RNA; RNA Interference; RNA-Binding Proteins; Rattus; Recommendation; Research; Research Personnel; Sampling; Seeds; Serine; Solid; Structure; Technology; Therapeutic; Time; TimeLine; Training; Transgenic Mice; age related neurodegeneration; career; cell type; experimental study; frontotemporal lobar dementia-amyotrophic lateral sclerosis; gene therapy; human monoclonal antibodies; in vivo; induced pluripotent stem cell; innovation; interdisciplinary approach; interest; molecular imaging; mouse model; multicatalytic endopeptidase complex; mutant; novel therapeutics; programs; protein TDP-43; protein aggregation; protein degradation; protein expression; protein purification; single molecule; superoxide dismutase 1; therapeutic gene; training opportunity; transcriptome sequencing; ubiquitin-protein ligase; Affinity; Alternative Splicing; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Antibodies; Antisense Oligonucleotides; Authorship; Award; Binding; Brain; Cell Line; Cell Nucleus; Cell model; Cells; Clinical Trials; Collaborations; Consensus; Cryo-electron tomography; Cryoelectron Microscopy; Cytoplasm; Cytoplasmic Protein; Dependovirus; Electron Microscopy; Environment; Excision; Failure; Foundations; Genes; Goals; Human; Immunoglobulin Fragments; In Situ; In Vitro; Lead; Letters; Liquid substance; Mediating; Mentors; Methods; Minor; Modeling; Molecular; Motor; Motor Neurons; Mus; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Nuclear; Nuclear RNA; Pathologic; Pathology; Pathway interactions; Phase; Physiological; Process; Productivity; Property; Proteins; Publications; RNA; RNA Interference; RNA-Binding Proteins; Rattus; Recommendation; Research; Research Personnel; Sampling; Seeds; Serine; Solid; Structure; Technology; Therapeutic; Time; TimeLine; Training; Transgenic Mice; age related neurodegeneration; career; cell type; experimental study; frontotemporal lobar dementia-amyotrophic lateral sclerosis; gene therapy; human monoclonal antibodies; in vivo; induced pluripotent stem cell; innovation; interdisciplinary approach; interest; molecular imaging; mouse model; multicatalytic endopeptidase complex; mutant; novel therapeutics; programs; protein TDP-43; protein aggregation; protein degradation; protein expression; protein purification; single molecule; superoxide dismutase 1; therapeutic gene; training opportunity; transcriptome sequencing; ubiquitin-protein ligase

In this new Pathway to Independence Award (K99/R00) application the candidate proposes to determine the structure and composition of phase separated TAR DNA-binding protein 43 (TDP-43) as a means to understand the pathology behind TDP-43 aggregates and to investigate approaches to ameliorate these aggregates. The candidate will acquire training in Cryo-EM methods, protein purification, single molecule imaging, and RNA sequencing through a combination of collaborations and coursework. The candidate, Dr. Yu, was regarded as an extremely promising junior investigator, with good productivity (including middle authorship on three publications since submission of this proposal), excellent previous training, and outstanding letters of recommendation. Reviewers agreed that the mentor, Dr. Cleveland, was excellent and remarked that he has an outstanding track record of mentoring many trainees who successfully transitioned to independent careers. The mentor also clearly stated that the candidate can take this research with him to establish his own research program. Collaborators provided strong letters of support and additional training opportunities. The environment was regarded as excellent, with many career and scientific opportunities available and a clear demonstration of institutional support for the candidate. Reviewers considered the training plan to be well-structured, appropriate to the candidate’s research and career goals, and further strengthened by inclusion of a detailed and clear timeline. The proposed research was regarded as exciting and highly innovative, with the potential to substantially impact the field. The research plan includes the use of state-of-the art methods and technologies and will provide a solid foundation from which the candidate can launch an independent career. Reviewers noted a few minor limitations with the research plan, including that it was not clear what samples would be used in the experiments. The research plan was also regarded as somewhat risky and potentially overly ambitious, but reviewers noted this was a negligible to minor concern given the demonstrated productivity of the candidate. In summary, the panel considered this an exciting proposal from an excellent candidate with only negligible to minor weaknesses that slightly lowered enthusiasm for some reviewers. As reflected in the final scores, most reviewers considered this an excellent proposal while others rated it as either outstanding or very good.

在这一新的独立奖项（K99/R00）应用中，候选提案确定相位分离的TAR DNA结合蛋白43（TDP-43）的结构和组成，作为了解TDP-43聚集体背后的病理和调查这些聚集物的方法的一种手段。候选人将通过合作和课程的组合进行冷冻EM方法，蛋白质纯化，单分子成像和RNA测序的培训。候选人Yu博士被认为是一位非常有前途的初级调查员，其生产力良好（包括自本提案提交以来三个出版物的中间作者资格），出色的先前培训和出色的推荐信。评论者同意，精神上的克利夫兰博士非常出色，并记得他在成功过渡到独立职业的许多学员心理记录中都有出色的记录。这位导师还清楚地指出，候选人可以与他一起进行这项研究以建立自己的研究计划。合作者提供了强有力的支持信和额外的培训机会。该环境被认为是极好的，拥有许多职业和科学机会，并明确证明了对候选人的机构支持。审稿人认为培训计划是结构良好的，适合候选人的研究和职业目标，并通过包括详细且清晰的时间表来进一步加强。拟议的研究被认为是令人兴奋和高度创新的，具有实质性影响该领域的潜力。该研究计划包括使用最先进的方法和技术，并将为候选人提供独立职业提供坚实的基础。审稿人指出了研究计划的一些小限制，包括尚不清楚实验中使用哪些样本。该研究计划也被认为是有些风险和可能过于雄心勃勃的，但审稿人指出，鉴于候选人的演示产物，这对微小的关注是可忽略的。总而言之，该小组认为这是一位出色的候选人的激动人心的提议，而这些候选人只能忽略一些小弱点，这些弱点略微降低了一些审稿人的热情。正如最终成绩所反映的那样，大多数审稿人认为这是一个很好的建议，而其他评论者则将其评为杰出或非常好。