Age Problem
Conditions
Keywords
helicase, induced pluripotent stem cells
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.
Interventions
OTHERtaking of cutaneous cells
Taking of cutaneous cells by biopsy Sample of blood
Sponsors
University Hospital, Montpellier
Study design
Observational model
OTHER
Time perspective
PROSPECTIVE
Eligibility
Sex/Gender
ALL
Age
18 Years to 65 Years
Healthy volunteers
No
Inclusion criteria
* Patinet with one of the 3 helicase-associated precoce aging desease
Exclusion criteria
* Minor and /or mentally incapable patient
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| 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
| Measure | Time frame | Description |
|---|---|---|
| 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) |
| 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. |
Countries
France
Outcome results
None listed