PMID- 31506453 OWN - NLM STAT- MEDLINE DCOM- 20201030 LR - 20210110 IS - 2045-2322 (Electronic) IS - 2045-2322 (Linking) VI - 9 IP - 1 DP - 2019 Sep 10 TI - Functional analysis of new human Bardet-Biedl syndrome loci specific variants in the zebrafish model. PG - 12936 LID - 10.1038/s41598-019-49217-7 [doi] LID - 12936 AB - The multiple genetic approaches available for molecular diagnosis of human diseases have made possible to identify an increasing number of pathogenic genetic changes, particularly with the advent of next generation sequencing (NGS) technologies. However, the main challenge lies in the interpretation of their functional impact, which has resulted in the widespread use of animal models. We describe here the functional modelling of seven BBS loci variants, most of them novel, in zebrafish embryos to validate their in silico prediction of pathogenicity. We show that target knockdown (KD) of known BBS (BBS1, BB5 or BBS6) loci leads to developmental defects commonly associated with ciliopathies, as previously described. These KD pleiotropic phenotypes were rescued by co-injecting human wild type (WT) loci sequence but not with the equivalent mutated mRNAs, providing evidence of the pathogenic effect of these BBS changes. Furthermore, direct assessment of cilia located in Kupffer's vesicle (KV) showed a reduction of ciliary length associated with all the studied variants, thus confirming a deleterious effect. Taken together, our results seem to prove the pathogenicity of the already classified and unclassified new BBS variants, as well as highlight the usefulness of zebrafish as an animal model for in vivo assays in human ciliopathies. FAU - Castro-Sanchez, Sheila AU - Castro-Sanchez S AD - Grupo de Biomarcadores Moleculares, Departamento de Bioquimica, Genetica e Inmunologia, Facultad de Biologia, Universidad de Vigo, Lagoas-Marcosende s/n, 36310, Vigo, Spain. shey.cs9@gmail.com. AD - Grupo de Investigacion en Enfermedades Raras y Medicina Pediatrica, Instituto de Investigacion Sanitaria Galicia Sur (IISGS), Vigo, Spain. shey.cs9@gmail.com. AD - Centro de Investigaciones Biomedicas (CINBIO), Centro Singular de Investigacion de Galicia 2016-2019, Universidad de Vigo, Vigo, Spain. shey.cs9@gmail.com. FAU - Suarez-Bregua, Paula AU - Suarez-Bregua P AD - Department of Biotechnology and Aquaculture, Institute of Marine Research, Spanish National Research Council (IIM-CSIC), Vigo, Spain. FAU - Novas, Rossina AU - Novas R AD - Human Molecular Genetics Laboratory, Institut Pasteur de Montevideo, Mataojo 2020, Montevideo, CP11400, Uruguay. FAU - Alvarez-Satta, Maria AU - Alvarez-Satta M AD - Grupo de Biomarcadores Moleculares, Departamento de Bioquimica, Genetica e Inmunologia, Facultad de Biologia, Universidad de Vigo, Lagoas-Marcosende s/n, 36310, Vigo, Spain. AD - Grupo de Investigacion en Enfermedades Raras y Medicina Pediatrica, Instituto de Investigacion Sanitaria Galicia Sur (IISGS), Vigo, Spain. AD - Centro de Investigaciones Biomedicas (CINBIO), Centro Singular de Investigacion de Galicia 2016-2019, Universidad de Vigo, Vigo, Spain. FAU - Badano, Jose L AU - Badano JL AUID- ORCID: 0000-0002-0706-8652 AD - Human Molecular Genetics Laboratory, Institut Pasteur de Montevideo, Mataojo 2020, Montevideo, CP11400, Uruguay. FAU - Rotllant, Josep AU - Rotllant J AD - Department of Biotechnology and Aquaculture, Institute of Marine Research, Spanish National Research Council (IIM-CSIC), Vigo, Spain. FAU - Valverde, Diana AU - Valverde D AD - Grupo de Biomarcadores Moleculares, Departamento de Bioquimica, Genetica e Inmunologia, Facultad de Biologia, Universidad de Vigo, Lagoas-Marcosende s/n, 36310, Vigo, Spain. dianaval@uvigo.es. AD - Grupo de Investigacion en Enfermedades Raras y Medicina Pediatrica, Instituto de Investigacion Sanitaria Galicia Sur (IISGS), Vigo, Spain. dianaval@uvigo.es. AD - Centro de Investigaciones Biomedicas (CINBIO), Centro Singular de Investigacion de Galicia 2016-2019, Universidad de Vigo, Vigo, Spain. dianaval@uvigo.es. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190910 PL - England TA - Sci Rep JT - Scientific reports JID - 101563288 RN - 0 (BBS5 protein, human) RN - 0 (Bbs1 protein, human) RN - 0 (Cytoskeletal Proteins) RN - 0 (MKKS protein, human) RN - 0 (Microtubule-Associated Proteins) RN - 0 (Oligonucleotides, Antisense) RN - 0 (Phosphate-Binding Proteins) RN - EC 3.6.1.- (Group II Chaperonins) SB - IM EIN - Sci Rep. 2020 Feb 13;10(1):2876. PMID: 32051508 MH - Animals MH - Bardet-Biedl Syndrome/genetics/*pathology MH - Cohort Studies MH - Cytoskeletal Proteins/antagonists & inhibitors/genetics/*metabolism MH - Disease Models, Animal MH - Embryo, Nonmammalian/metabolism/*pathology MH - Female MH - *Genetic Loci MH - Group II Chaperonins/antagonists & inhibitors/genetics/*metabolism MH - High-Throughput Nucleotide Sequencing MH - Humans MH - Male MH - Microtubule-Associated Proteins/antagonists & inhibitors/genetics/*metabolism MH - *Mutation MH - Oligonucleotides, Antisense/administration & dosage MH - Pedigree MH - Phenotype MH - Phosphate-Binding Proteins/antagonists & inhibitors/genetics/*metabolism MH - Zebrafish PMC - PMC6736949 COIS- The authors declare no competing interests. EDAT- 2019/09/12 06:00 MHDA- 2020/10/31 06:00 PMCR- 2019/09/10 CRDT- 2019/09/12 06:00 PHST- 2018/11/28 00:00 [received] PHST- 2019/08/16 00:00 [accepted] PHST- 2019/09/12 06:00 [entrez] PHST- 2019/09/12 06:00 [pubmed] PHST- 2020/10/31 06:00 [medline] PHST- 2019/09/10 00:00 [pmc-release] AID - 10.1038/s41598-019-49217-7 [pii] AID - 49217 [pii] AID - 10.1038/s41598-019-49217-7 [doi] PST - epublish SO - Sci Rep. 2019 Sep 10;9(1):12936. doi: 10.1038/s41598-019-49217-7.