Detection of cryptic exons (splicing differences) in TDP-43 as biomarkers for frontotemporal dementia and Alzheimer's disease

检测 TDP-43 中的隐秘外显子（剪接差异）作为额颞叶痴呆和阿尔茨海默病的生物标志物

• 批准号: 10913106
• 负责人: Andrew Singleton
• 金额: $ 478.57万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10913106
• 关键词: ALS patients; Address; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Biological Assay; Biological Markers; Central Nervous System; Cerebrospinal Fluid; Clinical; Deposition; Detection; Disease; Exons; Frontotemporal Dementia; Goals; Human; Induced pluripotent stem cell derived neurons; Lead; London; Manuscripts; Mass Spectrum Analysis; Monoclonal Antibodies; Nature; Neurons; Pathology; Patients; Peptides; Plasma; Printing; Progressive Supranuclear Palsy; Proteins; PubMed; RNA; RNA Splicing; Risk; Sampling; Specificity; Transcript; Universities; case control; clinical application; college; experimental study; frontotemporal lobar dementia amyotrophic lateral sclerosis; human tissue; induced pluripotent stem cell; industry partner; novel; polyclonal antibody; protein TDP-43; protein biomarkers; two-dimensional

After successfully addressing goals 1 and 2, we have been working towards identifying the main cryptic exon candidates in human CSF and plasma samples. Over the last year, we obtained more than 200 samples of human CSF from patients with ALS, progressive supranuclear palsy (PSP), and control cases to determine if TDP-43 cryptic exons are present in diseased but not control patients. Our efforts using patient CSF have been critical in confirming and expanding on our findings in iPSC neurons. We detected 65 candidate cryptic exons and performed two-dimensional targeted mass spectrometry to identify lead cryptic peptide candidates. The manuscript detailing these experiments is currently in revision, and a pre-print on our findings has been deposited to Biorxiv. We are now attempting to utilize the cryptic peptides and cryptic exons identified in our previous study to develop peptide and RNA-based biomarkers and ultra-sensitive bioassays for clinical application. We are currently working with industry partner BioMarin, as well as Dr. Pietro Fratta from University College London, and Dr. David Walt from Harvard University to further investigate the creation of biomarkers. Brown, Anna-Leigh, et al. "TDP-43 loss and ALS-risk SNPs drive mis-splicing and depletion of UNC13A." Nature 603.7899 (2022): 131-137. https://doi.org/10.1038/s41586-022-04436-3 PubMed ID 35197628 PubMed Central ID PMC8891020 Seddighi, Sahba, et al. Mis-spliced transcripts generate de novo proteins in TDP-43-related ALS/FTD (2023, submitted). Preprint via bioRxiv https://doi.org/10.1101/2023.01.23.525149

在成功解决目标1和2之后，我们一直致力于鉴定人CSF和血浆样品中的主要隐藏外显子候选物。在过去的一年里，我们从ALS、进行性核上性麻痹（PSP）和对照病例中获得了200多份人CSF样本，以确定TDP-43隐藏外显子是否存在于患病患者而非对照患者中。我们使用患者CSF的努力对于确认和扩展我们在iPSC神经元中的发现至关重要。我们检测了65个候选隐藏外显子，并进行了二维靶向质谱法，以确定潜在的潜在肽候选人。详细描述这些实验的手稿目前正在修订中，我们发现的预印本已存入Biorxiv。我们现在正试图利用我们以前的研究中发现的隐藏肽和隐藏外显子，开发基于肽和RNA的生物标志物和超灵敏的生物测定方法用于临床应用。我们目前正在与行业合作伙伴BioMarin以及伦敦大学学院的Pietro Fratta博士和哈佛大学的大卫沃尔特博士合作，进一步研究生物标志物的产生。 Brown，Anna-Leigh等人，“TDP-43损失和ALS风险SNP驱动UNC 13 A的错误剪接和耗尽。“Nature 603.7899（2022）：131-137. https://doi.org/10.1038/s41586-022-04436-3 PubMed ID 35197628 PubMed Central ID PMC8891020 Seddighi，Sahba，et al. Mis-spliced transcripts generate de novo proteins in TDP-43-related ALS/FTD（2023，submitted）.通过bioRxiv预印本https://doi.org/10.1101/2023.01.23.525149