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

在成功地解决了目标1和目标2之后，我们一直致力于在人脑脊液和血浆样本中确定主要的隐外显子候选者。在过去的一年里，我们从ALS患者、进行性核上性麻痹（PSP）患者和对照病例中获得了200多份人类脑脊液样本，以确定TDP-43隐外显子是否存在于患病患者而不存在于对照患者中。我们使用患者脑脊液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个候选的隐外显子，并进行了二维靶向质谱分析，以确定主要的隐肽候选。详细描述这些实验的手稿目前正在修订中，我们的研究结果的预印本已存入Biorxiv。我们现在正试图利用我们之前的研究中发现的隐肽和隐外显子来开发基于肽和rna的生物标志物和超灵敏的临床应用生物测定。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David Walt博士合作，进一步研究生物标志物的创造。

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