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Identification of TRP Channels as New Potential Therapeutic Targets in Primary and Secondary Raynaud's Phenomenon. (TRP)

Primary Purpose

Raynaud's Phenomenon Isolated Primary, Raynaud's Phenomenon (Secondary), Healthy

Status
Completed
Phase
Not Applicable
Locations
France
Study Type
Interventional
Intervention
Biopsy
Sponsored by
University Hospital, Grenoble
About
Eligibility
Locations
Arms
Outcomes
Full info

About this trial

This is an interventional basic science trial for Raynaud's Phenomenon Isolated Primary focused on measuring Transient Receptor Potential Channels, Raynaud's Phenomenon Isolated Primary, Raynaud's Phenomenon (Secondary)

Eligibility Criteria

18 Years - undefined (Adult, Older Adult)All SexesAccepts Healthy Volunteers

Inclusion Criteria:

  • Healthy volunteers :dated and signed written consent
  • Primary Raynaud's patients :
  • dated and signed written consent
  • subject presenting a primary Raynaud's phenomenon non associated to a connectivite. RP crisis are defined by a succession of caracteristic symptoms : a syncopal phase with distal whitening of the fingers, usually followed by a cyanotic phase, and then a hyperhemic phase with the outbreak of painful redness.The syncopal phase is always present, while the two other phases can be less pronounced, or even absent.
  • Sclerodermy secondary Raynaud's patients :
  • dated and signed written consent
  • subject with Raynaud's phenomenon secondary to systemic sclerosis, according to the 2013 EULAR/ACR criteria.

Exclusion Criteria:

  • Minor person or major person protected by the law
  • Exclusion period for any clinical trial including a medication or focusing on a clinical device linked to the pathology
  • Social secutrity scheme non-affiliated person or person who cannot pretend to such a scheme
  • Pregnant woman, parturient or breast-feeding mother
  • Person displaying a digital ulcer or a history of digital ulcer
  • Severe concurrent disease found at oral questioning : evolutive cancer, hepatic insufficiency, history of myocardic infarction in the past five years, angina pectoris
  • Hemodynamic instability
  • Any tobacco consumption
  • Any contraindication to the medication used in this trial :
  • Anesderm ® (lidocain/procain) : Known hypersensitivity, to local amide linking anesthetic or to any other compound of the cream, congenital methemoglobin porphyria
  • Lidocain Aguettant 5 mg/ml without preservative, injectable solution : Known hypersensitivity to lidocain chlorhydrate, to local amide linking anesthetic or to one of its excipients, patients affected with reccurrent porphyrias, administration using intravenous way for children under 5 years
  • Xylocain 10 mg/ml epinephrine 0.005 mg/ml, injectable solution : Known hypersensitivity to lidocain chlorhydrate, to local amide linking anesthetic or to one of its excipients, patients affected with reccurrent porphyrias, administration using intravenous way for children under 5 years, linked to the epinephrined compound : intra vascular way, angina pectoris, ventricular rythm troubles, severe arterial hypertension, obstructive cardiomyopathy, hyper thyroidy, anesthesia with local infiltration on the extermities (fingers, penis).
  • Kalinox ® (MEOPA) : Need for ventilation in pure oxygen, intracranial hypertension, alteration of consciousness, cranial trauma, pneumothorax, emphysema bubble, submarine diving accident, intestinal gas distension, gas bubble from an eye surgery under 3 months, known and non substituted B12 vitamine or folic acid deficiency, neurological disorder of recent onset and non explained.
  • Person deprived from his/her freedom on a judicial or administrative decision, person under a legal protection measure.

Sites / Locations

  • CHU of Grenoble Alpes

Arms of the Study

Arm 1

Arm 2

Arm 3

Arm Type

Experimental

Experimental

Experimental

Arm Label

Healthy

Primary Raynaud's phenomenon

Secondary Raynaud's phenomenon

Arm Description

Two sequences of ten healthy volunteers each will be performed : During the first sequence, a biopsy of the gluteal area and another of the surrounding nail area of a finger will be sampled.Then, the cutaneous blood flux of the hand will be recorded at rest and during cooling and warming periods. During the second sequence, two other biopsies of the surrounding nail area of two other fingers will be sampled.

Two sequences of ten primary Raynaud's syndrome each will be performed : During the first sequence, a biopsy of the gluteal area and another of the surrounding nail area of a finger will be sampled.Then, the cutaneous blood flux of the hand will be recorded at rest and during cooling and warming periods. During the second sequence, two other biopsies of the surrounding nail area of two other fingers will be sampled.

Two sequences of ten patients each will be performed : During the first sequence, a biopsy of the gluteal area and another of the surrounding nail area of a finger will be sampled.Then, the cutaneous blood flux of the hand will be recorded at rest and during cooling and warming periods. During the second sequence, two other biopsies of the surrounding nail area of two other fingers will be sampled.

Outcomes

Primary Outcome Measures

Comparison of the expression of mRNA of different TRP channels between the three groups.
Quantification of the expression of mRNA of TRP channels by RT PCR

Secondary Outcome Measures

Comparison of the expression of mRNA of different TRP channels between the three groups after a loco regional cooling.
RT PCR
Correlation between the expression of mRNA of different TRP channels and the cutaneous blood flux between the three groups during a loco regional cooling.
RT PCR, laser speckle contrast imaging, infra red thermogrphy, tissue viability imaging
Correlation between the expression of mRNA of different TRP channels and the cutaneous blood flux between the three groups during a loco regional warming at 39°C and 43°C.
RT PCR, laser doppler
Comparison of the expression of mRNA of different TRP channels between the biopsies of an affected area (surrouding nail area) and a non-affected area (gluteal region) in the three groups.
RT PCR
Comparison of expressed micro RNA between the three groups.
RT PCR
Comparison of the methylation states of CpG clusters found at the level of dysregulated genes.
RT PCR
Comparison of the proportion of myo fibroblasts with or without pharmacological modulation on the TRP(s) which expression is modified in RP.
Immuno fluorescence with an anti-alpha smooth muscle cell antibody

Full Information

First Posted
June 20, 2017
Last Updated
April 8, 2020
Sponsor
University Hospital, Grenoble
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1. Study Identification

Unique Protocol Identification Number
NCT03211325
Brief Title
Identification of TRP Channels as New Potential Therapeutic Targets in Primary and Secondary Raynaud's Phenomenon.
Acronym
TRP
Official Title
Identification of Transient Receptor Potential (TRP) Channels as New Potential Therapeutic Targets in Primary and Secondary Raynaud's Phenomenon.
Study Type
Interventional

2. Study Status

Record Verification Date
April 2019
Overall Recruitment Status
Completed
Study Start Date
October 18, 2016 (Actual)
Primary Completion Date
March 17, 2019 (Actual)
Study Completion Date
June 27, 2019 (Actual)

3. Sponsor/Collaborators

Responsible Party, by Official Title
Sponsor
Name of the Sponsor
University Hospital, Grenoble

4. Oversight

Studies a U.S. FDA-regulated Drug Product
No
Studies a U.S. FDA-regulated Device Product
No
Product Manufactured in and Exported from the U.S.
No
Data Monitoring Committee
Yes

5. Study Description

Brief Summary
The investigators seek to perform a comparative transcriptome analysis of TRP channels residing in the skin between primary and scleroderma secondary Raynaud's phenomenon and healthy subjects.
Detailed Description
Raynaud's phenomenon (RP) is a clinical syndrome that results from an excessive vascular reactivity of the microcirculation of the fingers and/or other extremities of the body in response to a stress, mostly cold. Primary RP affects 4 to 6% of the general population and displays important geographic variations. Most secondary RP are linked to systemic scleroderma, a rare condition affecting mainly women. The systemic scleroderma is characterized by a cutaneous and visceral fibrosis and is associated with micro vascular damages and the production of self antibodies directed against specific cellular antigenes. The American College of Rheumatology in association with the European League Against Rheumatism has recently set diagnostic criteria that define this disease more precisely. Besides, clinical forms without cutaneous infringment have been described as limited scleroderma. Cutaneous limited forms are characterized by cutaneous fibrosis affecting mainly hands, arms and face. RP is the first clinical sign of these forms ; anti-centromer antibodies are found in 50 to 90% patients. Diffuse cutaneous scleroderma is a fast progressing disease that affects most parts of the skin and internal organs and displays the presence of anti topoisomerase antibodies in 30% of the patients. Whether it is primary or secondary, severe forms of RP can result in an important functional disability. Besides that, iterative ischemic episodes contribute to the onset of trophic disorders that are associated with a higher morbidity. These trophic disorders are mainly encountered in the most severe forms, especially those that are secondary to systemic scleroderma ; a third of which will turn into gangrene or amputation, leading to an important disability for these patients. Up to now, no curative treatment of these ulcers has displayed a convincing efficacy. Despite its frequency and the potential seriousness of its complications, therapeutic care options for RP remain limited. Conservative measures are recommended in the first place, especially avoiding cold exposure and wearing warm clothes. When those measures do not suffice, pharmacological treatment can be introduced. Only calcic inhibitors are authorized for this indication but their effect on the frequency of the crises is modest (reduction of 2.8 crises per week on average). In scleroderma secondary RP, the effect appears to be modest as well according to the severity of the affection and displays a reduction of 4 crises per week on average. Besides that, this kind of treatment, when taken continuously, exposes the patients to side effects (inferior limb edema in particular). Other vasodilators such as type 5 phosphodiesterase have a significant effect, but the latter remains low in secondary RPs. Finally, intravenous injections of Iloprost on a sequential scheme have displayed a superior effect compared to Nifedipine, but result in intense and frequent side effects that limit considerably the use of this medication for a common practice. Therefore, there is a real need for new therapeutic targets in RP and this requires a better knowledge of the physiopathology of this phenomenon. Whether it is primary or secondary, microvascular dysfunction is now considered as a key element in the physiopathogeny of RP. Some ionic channels from the Transient Receptor Potential Channels' family react to temperature variations and are therefore involved in the microvascular response. In the cutaneous tissue, multiple TRP channels are present on the afferent neurons and are involved in the response to hot temperatures (especially TRPV1, TRPV3 and TRPV4) or to environmental cold (TRPM8) or hyper algic cold (TRPA1). Although they were initially reduced to an afferent role towards thermic or chemical stimuli, TRP channels have been lately recognized as having more complex functions, especially in the cutaneous homeostasis sustainibility, the growth and cellular differentiation and even the inflammatory response. Thus, the recent identification of numerous TRP channels (TRPV1, TRPV3, TRPV4, TRPM8, TRPA1, TRPC1, TRPC4, TRPV6) on the surface of keratinocytes suggests their involvement in the differenciation of these cells. Although their precise role and barrier function in the skin remain unknown, anomalies of the expression of some TRP channels have been associated with specific dermatologic diseases, and have recently being the issue of a review. Nevertheless, TRP expression in Raynaud's phenomenon has been investigated only few times in human skin . Considering the major role of the micro vascular dysfunction in RP and the involvement of specific TRP channels in the regulation of the cutaneous microvascular tone, the investigators put forward the hypothesis that the expression of some of these channels could be modified in primary and scleroderma secondary RP forms. This hypothesis is supported by an experimental trial (on mouse model of systemic cutaneous scleroderma), in which the activation of TRPV1 has been associated to the release of vasodilator peptides that displayed a protective effect against fibrosis and thus suggesting a plausible role of TRPV1 in the physiopathology of cutaneous systemic scleroderma. Although RP strikes different parts of the body (hands, feet, ears...), the essential topography of Raynaud's crises remains hands. In scleroderma secondary RP, the microcirculatory damages that are the easiest to reach are the areas surrounding nails. The latters can be examined using a non-invasive technique that is capillaroscopy and sometimes even to the naked eye. Histology by cutaneous biopsy can be made as well. The investigators propose here to sample cutaneous biopsies in these areas, that are on the one hand the most pathognomonic to display the microvascular damages and on the other hand, concerned by the Raynaud's crises. Now studied as potential therapeutic targets for many disorders, TRP channels could represent a very interesting track in severe RP forms. The current development of numerous agonists and inhibitors of TRP channels could allow to foresee a clinical trial in Raynaud's phenomenon. On a physiopathological point of view, a better knowledge of the potential role of TRP channels in this disease could also explain the differences that have been observed between primary and secondary Raynaud's phenomenons.

6. Conditions and Keywords

Primary Disease or Condition Being Studied in the Trial, or the Focus of the Study
Raynaud's Phenomenon Isolated Primary, Raynaud's Phenomenon (Secondary), Healthy
Keywords
Transient Receptor Potential Channels, Raynaud's Phenomenon Isolated Primary, Raynaud's Phenomenon (Secondary)

7. Study Design

Primary Purpose
Basic Science
Study Phase
Not Applicable
Interventional Study Model
Parallel Assignment
Masking
None (Open Label)
Allocation
Non-Randomized
Enrollment
29 (Actual)

8. Arms, Groups, and Interventions

Arm Title
Healthy
Arm Type
Experimental
Arm Description
Two sequences of ten healthy volunteers each will be performed : During the first sequence, a biopsy of the gluteal area and another of the surrounding nail area of a finger will be sampled.Then, the cutaneous blood flux of the hand will be recorded at rest and during cooling and warming periods. During the second sequence, two other biopsies of the surrounding nail area of two other fingers will be sampled.
Arm Title
Primary Raynaud's phenomenon
Arm Type
Experimental
Arm Description
Two sequences of ten primary Raynaud's syndrome each will be performed : During the first sequence, a biopsy of the gluteal area and another of the surrounding nail area of a finger will be sampled.Then, the cutaneous blood flux of the hand will be recorded at rest and during cooling and warming periods. During the second sequence, two other biopsies of the surrounding nail area of two other fingers will be sampled.
Arm Title
Secondary Raynaud's phenomenon
Arm Type
Experimental
Arm Description
Two sequences of ten patients each will be performed : During the first sequence, a biopsy of the gluteal area and another of the surrounding nail area of a finger will be sampled.Then, the cutaneous blood flux of the hand will be recorded at rest and during cooling and warming periods. During the second sequence, two other biopsies of the surrounding nail area of two other fingers will be sampled.
Intervention Type
Procedure
Intervention Name(s)
Biopsy
Primary Outcome Measure Information:
Title
Comparison of the expression of mRNA of different TRP channels between the three groups.
Description
Quantification of the expression of mRNA of TRP channels by RT PCR
Time Frame
at the moment of the first biopsy
Secondary Outcome Measure Information:
Title
Comparison of the expression of mRNA of different TRP channels between the three groups after a loco regional cooling.
Description
RT PCR
Time Frame
after cooling (2hours after the first biopsy)
Title
Correlation between the expression of mRNA of different TRP channels and the cutaneous blood flux between the three groups during a loco regional cooling.
Description
RT PCR, laser speckle contrast imaging, infra red thermogrphy, tissue viability imaging
Time Frame
after cooling (2hours after the first biopsy)
Title
Correlation between the expression of mRNA of different TRP channels and the cutaneous blood flux between the three groups during a loco regional warming at 39°C and 43°C.
Description
RT PCR, laser doppler
Time Frame
after warming (3 hours after the first biopsy)
Title
Comparison of the expression of mRNA of different TRP channels between the biopsies of an affected area (surrouding nail area) and a non-affected area (gluteal region) in the three groups.
Description
RT PCR
Time Frame
at the moment of the biopsies
Title
Comparison of expressed micro RNA between the three groups.
Description
RT PCR
Time Frame
at the moment of the first biopsy
Title
Comparison of the methylation states of CpG clusters found at the level of dysregulated genes.
Description
RT PCR
Time Frame
at the moment of the first biopsy
Title
Comparison of the proportion of myo fibroblasts with or without pharmacological modulation on the TRP(s) which expression is modified in RP.
Description
Immuno fluorescence with an anti-alpha smooth muscle cell antibody
Time Frame
second sequence (The interval between the two sequence must not exceed 180 days)

10. Eligibility

Sex
All
Minimum Age & Unit of Time
18 Years
Accepts Healthy Volunteers
Accepts Healthy Volunteers
Eligibility Criteria
Inclusion Criteria: Healthy volunteers :dated and signed written consent Primary Raynaud's patients : dated and signed written consent subject presenting a primary Raynaud's phenomenon non associated to a connectivite. RP crisis are defined by a succession of caracteristic symptoms : a syncopal phase with distal whitening of the fingers, usually followed by a cyanotic phase, and then a hyperhemic phase with the outbreak of painful redness.The syncopal phase is always present, while the two other phases can be less pronounced, or even absent. Sclerodermy secondary Raynaud's patients : dated and signed written consent subject with Raynaud's phenomenon secondary to systemic sclerosis, according to the 2013 EULAR/ACR criteria. Exclusion Criteria: Minor person or major person protected by the law Exclusion period for any clinical trial including a medication or focusing on a clinical device linked to the pathology Social secutrity scheme non-affiliated person or person who cannot pretend to such a scheme Pregnant woman, parturient or breast-feeding mother Person displaying a digital ulcer or a history of digital ulcer Severe concurrent disease found at oral questioning : evolutive cancer, hepatic insufficiency, history of myocardic infarction in the past five years, angina pectoris Hemodynamic instability Any tobacco consumption Any contraindication to the medication used in this trial : Anesderm ® (lidocain/procain) : Known hypersensitivity, to local amide linking anesthetic or to any other compound of the cream, congenital methemoglobin porphyria Lidocain Aguettant 5 mg/ml without preservative, injectable solution : Known hypersensitivity to lidocain chlorhydrate, to local amide linking anesthetic or to one of its excipients, patients affected with reccurrent porphyrias, administration using intravenous way for children under 5 years Xylocain 10 mg/ml epinephrine 0.005 mg/ml, injectable solution : Known hypersensitivity to lidocain chlorhydrate, to local amide linking anesthetic or to one of its excipients, patients affected with reccurrent porphyrias, administration using intravenous way for children under 5 years, linked to the epinephrined compound : intra vascular way, angina pectoris, ventricular rythm troubles, severe arterial hypertension, obstructive cardiomyopathy, hyper thyroidy, anesthesia with local infiltration on the extermities (fingers, penis). Kalinox ® (MEOPA) : Need for ventilation in pure oxygen, intracranial hypertension, alteration of consciousness, cranial trauma, pneumothorax, emphysema bubble, submarine diving accident, intestinal gas distension, gas bubble from an eye surgery under 3 months, known and non substituted B12 vitamine or folic acid deficiency, neurological disorder of recent onset and non explained. Person deprived from his/her freedom on a judicial or administrative decision, person under a legal protection measure.
Overall Study Officials:
First Name & Middle Initial & Last Name & Degree
Sophie Blaise, MD, PhD
Organizational Affiliation
University Hospital, Grenoble
Official's Role
Principal Investigator
Facility Information:
Facility Name
CHU of Grenoble Alpes
City
Grenoble
State/Province
Isère
ZIP/Postal Code
38700
Country
France

12. IPD Sharing Statement

Plan to Share IPD
Undecided
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Identification of TRP Channels as New Potential Therapeutic Targets in Primary and Secondary Raynaud's Phenomenon.

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