Pathology of Helicases and Premature Aging: Study by Derivation of hiPS
- Conditions
- Age Problem
- Interventions
- Other: taking of cutaneous cells
- Registration Number
- NCT03898817
- Lead Sponsor
- University Hospital, Montpellier
- Brief Summary
Topic of this work is the involvement of replicative helicases in human premature ageing syndrome. Replicative helicases are ubiquitous and essential during numerous reactions of the DNA metabolism.
The family of replicative helicases (RecQL) is involved in the replication/repair of the DNA and in the telomere maintenance. There are 5 enzymes in human and 3 of them are involved in clinically recognizable syndromes: WRN for the Werner syndrome, BLM for the Bloom syndrome and RECQL4 for the Rothmund Thomson syndrome. All are responsive of a high cancer risk due to genomic instability. Molecular and cellular mechanisms involved in these diseases of ageing are unknown. Moreover, for all of them, there is not therapeutic or preventive solution.
- Detailed Description
For understanding the involved mechanisms we would like to model the 3 diseases with hiPS (human induced Pluripotent Stem cells) from somatic cells of patients. The patient recruitment was organized by the Montpellier and Nîmes public hospitals.
The project is to generate a hiPS cell line for the 3 syndromes from fibroblasts and/or blood samples. Then, we could induce differentiation of hiPS to a target cell line of the diseases. Finally we could study the disease development following the genomic instability (karyotype, array-CGH) and the cellular ageing (senescence-associated heterochromatin foci, telomere length).
For each mutated enzyme, we will perform a transcriptional profiling (splice, mRNA quantification) and protein studies (western blot). All results will be compared to wild type cells.
Recruitment & Eligibility
- Status
- TERMINATED
- Sex
- All
- Target Recruitment
- 3
- Patinet with one of the 3 helicase-associated precoce aging desease
- Minor and /or mentally incapable patient
Study & Design
- Study Type
- OBSERVATIONAL
- Study Design
- Not specified
- Arm && Interventions
Group Intervention Description Taking of cutaneous cells by biopsy taking of cutaneous cells Taking of cutaneous cells by biopsy and a sample of blood
- Primary Outcome Measures
Name Time Method Genomic instability : analysis 1 year Molecular analysis of hiPS cell derived from pathological tissue (karyotype, array-CGH)
Genomic instability : size of telomers 1 year size of the telomers which will be quantified under microscope after fluorescent marking in situ of telomeric sequences (Q-FISH technique)
Genomic instability : Duplication of centrosomes 1 year duplication of centrosomes which is often associated with chromosomal segregation errors and genomic instability. This analysis will be done by immunolabelling using antibodies specific to the 2 main components of centrosomes, pericentrin and -tubulin.
- Secondary Outcome Measures
Name Time Method cellular ageing : molecular characterization 2 years lengthening of telomeric sequence size (Q-FISH), re-expression of pluripotency genes (QRTPCR), transcriptional profile of iPS cell lines.
cellular ageing : molecular analysis of hiPS cell derived from pathological tissue 2 years Analysis of senescence-associated heterochromatin foci, telomere length (Q-FISH)
cellular ageing : IPS line with the criteria defined for morphological characterization 2 years expression of specific surface markers (specifics markers : TRA-1-60, SSEA-4), ability to re-differentiate in the 3 embryonic layers (specifics markers : SMA, MAP2, FOXA2)
Trial Locations
- Locations (1)
University Hospital Montpellier
🇫🇷Montpellier, France