Pathology of Helicases and Premature Aging: Study by Derivation of hiPS (HeliPS)
|The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Read our disclaimer for details.|
|ClinicalTrials.gov Identifier: NCT03898817|
Recruitment Status : Completed
First Posted : April 2, 2019
Last Update Posted : May 1, 2019
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.
|Condition or disease||Intervention/treatment|
|Age Problem||Other: taking of cutaneous cells|
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.
|Study Type :||Observational|
|Actual Enrollment :||3 participants|
|Official Title:||Pathology of Helicases and Premature Aging: Study by Derivation of hiPS|
|Study Start Date :||September 2015|
|Actual Primary Completion Date :||September 2017|
|Actual Study Completion Date :||March 2019|
Taking of cutaneous cells by biopsy
Taking of cutaneous cells by biopsy and a sample of blood
Other: taking of cutaneous cells
Taking of cutaneous cells by biopsy Sample of blood
- Genomic instability : analysis [ Time Frame: 1 year ]Molecular analysis of hiPS cell derived from pathological tissue (karyotype, array-CGH)
- Genomic instability : size of telomers [ Time Frame: 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 [ Time Frame: 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.
- cellular ageing : molecular analysis of hiPS cell derived from pathological tissue [ Time Frame: 2 years ]Analysis of senescence-associated heterochromatin foci, telomere length (Q-FISH)
- cellular ageing : IPS line with the criteria defined for morphological characterization [ Time Frame: 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 [ Time Frame: 2 years ]lengthening of telomeric sequence size (Q-FISH), re-expression of pluripotency genes (QRTPCR), transcriptional profile of iPS cell lines.
To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.
Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT03898817
|University Hospital Montpellier|
|Montpellier, France, 34000|
|Principal Investigator:||Vincent GATINOIS, harmD||University Hospital, Montpellier|