Pathology of Helicases and Premature Aging: Study by Derivation of hiPS (HeliPS)

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.

Study Overview

• Status: Terminated
• Conditions: Age Problem
• Intervention / Treatment: Other: taking of cutaneous cells

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.

• Study Type: Observational
• Enrollment (Actual): 3

Contacts and Locations

Study Locations

• France
  • Montpellier, France, 34000
    • University Hospital Montpellier

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 65 years (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

• Sampling Method: Probability Sample

Study Population

Population with pathology of helicases and premature aging

Description

Inclusion Criteria:

• Patinet with one of the 3 helicase-associated precoce aging desease

Exclusion Criteria:

• Minor and /or mentally incapable patient

Study Plan

How is the study designed?

Design Details

• Observational Models: Other
• Time Perspectives: Prospective

Number of groups / cohorts

1

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
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

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Genomic instability : analysis	Molecular analysis of hiPS cell derived from pathological tissue (karyotype, array-CGH)	1 year
Genomic instability : size of telomers	size of the telomers which will be quantified under microscope after fluorescent marking in situ of telomeric sequences (Q-FISH technique)	1 year
Genomic instability : Duplication of centrosomes	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.	1 year

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
cellular ageing : molecular analysis of hiPS cell derived from pathological tissue	Analysis of senescence-associated heterochromatin foci, telomere length (Q-FISH)	2 years
cellular ageing : IPS line with the criteria defined for morphological characterization	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)	2 years
cellular ageing : molecular characterization	lengthening of telomeric sequence size (Q-FISH), re-expression of pluripotency genes (QRTPCR), transcriptional profile of iPS cell lines.	2 years

Collaborators and Investigators

• Sponsor: University Hospital, Montpellier
• Investigators: Principal Investigator: Vincent GATINOIS, harmD, University Hospital, Montpellier

Study record dates

Study Major Dates

• Study Start (Actual): September 9, 2015
• Primary Completion (Actual): September 9, 2017
• Study Completion (Actual): September 9, 2017

Study Registration Dates

• First Submitted: September 7, 2016
• First Submitted That Met QC Criteria: March 29, 2019
• First Posted (Actual): April 2, 2019

Study Record Updates

• Last Update Posted (Actual): December 29, 2021
• Last Update Submitted That Met QC Criteria: December 8, 2021
• Last Verified: December 1, 2021

More Information

Terms related to this study

• Keywords: helicase; induced pluripotent stem cells
• Additional Relevant MeSH Terms: Aging, Premature
• Other Study ID Numbers: 9360

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No