Periodic Reporting for period 4 - TREATCilia (Novel Therapeutic Avenues for dynein-related Ciliopathies)
Reporting period: 2021-08-01 to 2022-01-31
TREATCilia aimed to uncover underlying molecular pathophysiology in dynein based ciliopathies from gene to mechanism and we hope to identify future pharmacological entry points to ameliorate ciliary dynein-related phenotypes. Emplyoing exome sequencing, we were able to identify several novel human disease genes, such as PIH1D3, MNS1 and DNAH9, causing cilia motility defects and/or laterality defects when mutated as well as several novel genes associated with centra nervous system defects. In addition, we were able to establish a number of phenotype genotype correlation for motile ciliopathies/laterality disorders, inherited renal and skeletal phenotypes and neuro-developmental disorders By creating several of dynein-2- and IFT-mutant cell lines harboring human disease alleles using CRISPR base editing technologies, the project achieved generation of unique in-vitro ciliary condrodysplasia disease models. Multi-omics analyses of these model suggest disturbed Golgi-transport, actin-cytoskeleton modifications and changed extracellular matrix composition in mutants. We further identified a number of affected cell signalling pathways, offering putative future therapeutic targets.
With regards to non-motile ciliopathies, we focus on reno-skeletal disorders such as Jeune Syndrom and Short-Rib-Polydactyly Syndrome. In addition to gene identification and genotype-phenotype studies, we have recreated several hypomorphic patient alleles using CRISPR/Cas technologies and we are investigating downstream effects using Transcriptomics and Proteomics approaches. Chlamoydomonas drugscreening is currently prepared.