Tool for Fibromyalgia Diagnosis and Effect of Extra Virgin Olive Oil (FIBROKIT)

• ClinicalTrials.gov Identifier: NCT05921409
• Recruitment Status: Active, not recruiting
• First Posted: June 27, 2023
• Last Update Posted: July 14, 2023
• Sponsor: Pronacera Therapeutics SL
• Collaborator: Centre for the Development of Industrial Technology, Spain
• Information provided by (Responsible Party): Pronacera Therapeutics SL

Tracking Information

• First Submitted Date: June 19, 2023
• First Posted Date: June 27, 2023
• Last Update Posted Date: July 14, 2023
• Actual Study Start Date: January 1, 2021
• Estimated Primary Completion Date: August 31, 2023 (Final data collection date for primary outcome measure)

Current Primary Outcome Measures (submitted: June 27, 2023)

• Concentration of Voltage dependent anion channel 1 protein (VDAC) [ Time Frame: 0 weeks ]
  VDAC protein determination by western blot
• Concentration of Voltage dependent anion channel 1 protein (VDAC) [ Time Frame: 12 weeks ]
  VDAC protein determination by western blot
• Concentration of Voltage dependent anion channel 1 protein (VDAC) [ Time Frame: 24 weeks ]
  VDAC protein determination by western blot
• Concentration of Voltage dependent anion channel 1 protein (VDAC) [ Time Frame: 48 weeks ]
  VDAC protein determination by western blot
• Concentration of Microtubule-associated proteins 1B light chain 3B (LC3B) [ Time Frame: 0 weeks ]
  LC3B protein determination by western blot
• Concentration of Microtubule-associated proteins 1B light chain 3B (LC3B) [ Time Frame: 12 weeks ]
  LC3B protein determination by western blot
• Microtubule-associated proteins 1B light chain 3B (LC3B) [ Time Frame: 24 weeks ]
  LC3B protein determination by western blot
• Concentration of Microtubule-associated proteins 1B light chain 3B (LC3B) [ Time Frame: 48 weeks ]
  LC3B protein determination by western blot
• Concentration of Plasma Proteins [ Time Frame: 0 weeks ]
  Plasma proteome Analysis by label-free nano Liquid Chromatography coupled to tandem mass spectrometry after top-14 depletion
• Concentration of Plasma Proteins [ Time Frame: 12 weeks ]
  Plasma proteome Analysis by label-free nano Liquid Chromatography coupled to tandem mass spectrometry after top-14 depletion
• Concentration of Plasma Proteins [ Time Frame: 24 weeks ]
  Plasma proteome Analysis by label-free nano Liquid Chromatography coupled to tandem mass spectrometry after top-14 depletion
• Concentration of Plasma Proteins [ Time Frame: 48 weeks ]
  Plasma proteome Analysis by label-free nano Liquid Chromatography coupled to tandem mass spectrometry after top-14 depletion
• Abundance of Bacteria from the Intestinal Microbiome [ Time Frame: 0 weeks ]
  Fecal metagenomic analysis by amplicon sequencing of hypervariable region V3-V4 from RNA 16S bacterial gene
• Abundance of Bacteria from the Intestinal Microbiome [ Time Frame: 12 weeks ]
  Fecal metagenomic analysis by amplicon sequencing of hypervariable region V3-V4 from RNA 16S bacterial gene
• Abundance of Bacteria from the Intestinal Microbiome [ Time Frame: 24 weeks ]
  Fecal metagenomic analysis by amplicon sequencing of hypervariable region V3-V4 from RNA 16S bacterial gene
• Abundance of Bacteria from the Intestinal Microbiome [ Time Frame: 48 weeks ]
  Fecal metagenomic analysis by amplicon sequencing of hypervariable region V3-V4 from RNA 16S bacterial gene

Original Primary Outcome Measures (submitted: June 19, 2023)

• Voltage dependent anion channel 1 protein (VDAC) [ Time Frame: 0 weeks ]
  VDAC protein determination by western blot
• Voltage dependent anion channel 1 protein (VDAC) [ Time Frame: 12 weeks ]
  VDAC protein determination by western blot
• Voltage dependent anion channel 1 protein (VDAC) [ Time Frame: 24 weeks ]
  VDAC protein determination by western blot
• Voltage dependent anion channel 1 protein (VDAC) [ Time Frame: 48 weeks ]
  VDAC protein determination by western blot
• Microtubule-associated proteins 1B light chain 3B (LC3B) [ Time Frame: 0 weeks ]
  LC3B protein determination by western blot
• Microtubule-associated proteins 1B light chain 3B (LC3B) [ Time Frame: 12 weeks ]
  LC3B protein determination by western blot
• Microtubule-associated proteins 1B light chain 3B (LC3B) [ Time Frame: 24 weeks ]
  LC3B protein determination by western blot
• Microtubule-associated proteins 1B light chain 3B (LC3B) [ Time Frame: 48 weeks ]
  LC3B protein determination by western blot
• Plasma Proteome Analysis [ Time Frame: 0 weeks ]
  Plasma proteome Analysis by label-free nano Liquid Chromatography coupled to tandem mass spectrometry
• Plasma Proteome Analysis [ Time Frame: 12 weeks ]
  Plasma proteome Analysis by label-free nano Liquid Chromatography coupled to tandem mass spectrometry
• Plasma Proteome Analysis [ Time Frame: 24 weeks ]
  Plasma proteome Analysis by label-free nano Liquid Chromatography coupled to tandem mass spectrometry
• Plasma Proteome Analysis [ Time Frame: 48 weeks ]
  Plasma proteome Analysis by label-free nano Liquid Chromatography coupled to tandem mass spectrometry
• Intestinal Microbiome [ Time Frame: 0 weeks ]
  Fecal metagenomic analysis by amplicon sequencing of hypervariable region V3-V4 from RNA 16S bacterial gene
• Intestinal Microbiome [ Time Frame: 12 weeks ]
  Fecal metagenomic analysis by amplicon sequencing of hypervariable region V3-V4 from RNA 16S bacterial gene
• Intestinal Microbiome [ Time Frame: 24 weeks ]
  Fecal metagenomic analysis by amplicon sequencing of hypervariable region V3-V4 from RNA 16S bacterial gene
• Intestinal Microbiome [ Time Frame: 48 weeks ]
  Fecal metagenomic analysis by amplicon sequencing of hypervariable region V3-V4 from RNA 16S bacterial gene

Current Secondary Outcome Measures (submitted: June 27, 2023)

• 36-Item Short Form Health Survey (SF-36) Score [ Time Frame: 0 weeks ]
  36-Item Short Form Health Survey score
• 36-Item Short Form Health Survey (SF-36) Score [ Time Frame: 12 weeks ]
  36-Item Short Form Health Survey score
• 36-Item Short Form Health Survey (SF-36) Score [ Time Frame: 24 weeks ]
  36-Item Short Form Health Survey score
• 36-Item Short Form Health Survey (SF-36) Score [ Time Frame: 48 weeks ]
  36-Item Short Form Health Survey score
• Fibromyalgia Impact Questionnaire (FIQ) Score [ Time Frame: 0 weeks ]
  Fibromyalgia Impact Questionnaire (FIQ) score
• Fibromyalgia Impact Questionnaire (FIQ) Score [ Time Frame: 12 weeks ]
  Fibromyalgia Impact Questionnaire (FIQ) score
• Fibromyalgia Impact Questionnaire (FIQ) Score [ Time Frame: 24 weeks ]
  Fibromyalgia Impact Questionnaire (FIQ) score
• Fibromyalgia Impact Questionnaire (FIQ) Score [ Time Frame: 48 weeks ]
  Fibromyalgia Impact Questionnaire (FIQ) score

Original Secondary Outcome Measures (submitted: June 19, 2023)

• 36-Item Short Form Health Survey (SF-36) [ Time Frame: 0 weeks ]
  36-Item Short Form Health Survey score
• 36-Item Short Form Health Survey (SF-36) [ Time Frame: 12 weeks ]
  36-Item Short Form Health Survey score
• 36-Item Short Form Health Survey (SF-36) [ Time Frame: 24 weeks ]
  36-Item Short Form Health Survey score
• 36-Item Short Form Health Survey (SF-36) [ Time Frame: 48 weeks ]
  36-Item Short Form Health Survey score
• Fibromyalgia Impact Questionnaire (FIQ) [ Time Frame: 0 weeks ]
  Fibromyalgia Impact Questionnaire (FIQ) score
• Fibromyalgia Impact Questionnaire (FIQ) [ Time Frame: 12 weeks ]
  Fibromyalgia Impact Questionnaire (FIQ) score
• Fibromyalgia Impact Questionnaire (FIQ) [ Time Frame: 24 weeks ]
  Fibromyalgia Impact Questionnaire (FIQ) score
• Fibromyalgia Impact Questionnaire (FIQ) [ Time Frame: 48 weeks ]
  Fibromyalgia Impact Questionnaire (FIQ) score

• Current Other Pre-specified Outcome Measures: Not Provided
• Original Other Pre-specified Outcome Measures: Not Provided

Descriptive Information

• Brief Title: Tool for Fibromyalgia Diagnosis and Effect of Extra Virgin Olive Oil
• Official Title: Development of a New Tool for the Diagnosis and Monitoring of Fibromyalgia "FIBROKIT"

Brief Summary

PRONACERA THERAPEUTICS S.L. is a young biotechnological company focused on the development of genetic diagnostic tools and treatments for pathologies in different medical areas such as reproduction, fibromyalgia and rare diseases or with deficiencies in diagnosis, with the aim of helping to optimize health systems through improvement in terms and forms of diagnosis. Among the multiple lines of R+D+i that are currently being developed, the reproductive genomic line and endometrial functional molecular biology stand out. Specifically, it develops the design of markers and performs the genetic analysis of infertility focused on female endometrial tissue.

Fibromyalgia is a complex and common chronic pain disorder that affects 12 million Europeans. Along with other symptoms, fibromyalgia causes pain and general tenderness to touch. Currently this disease is diagnosed following the criteria established by the American College of Rheumatology (ACR) of a combination of relevant symptoms and the description of how the person feels. In other words, in practice it is a diagnosis by elimination. A patient suffering from fibromyalgia usually takes between 2 to 3 years to obtain the correct diagnosis. Health experts consider that fibromyalgia is a disease that is difficult to diagnose and that is associated with an expensive use of health system services.

With FIBROKIT, Pronacera aims to design and develop a new diagnostic and monitoring tool for fibromyalgia by designing a panel of specific plasma proteome and intestinal microbiome biomarkers and reducing the number of biological samples used. During the execution of this project, the company will have three leading research and innovation organizations in the sector (Helix BioS, CINUSA and CICbioGUNE) that will support Pronacera. FIBROKIT will have a cohort of 250 participants (206 patients and 44 healthy volunteers) to validate the diagnostic capacity of the tool and perform a robust biostatistical study.

Detailed Description

Fibromyalgia (FM) is a chronic syndrome characterized mainly by the presence of widespread pain. This pain is usually accompanied by other alterations, both physically and psychologically, which, ultimately, seriously compromise people's quality of life, affecting their family, social and work environment and causing high social-sanitary costs. Although different factors such as an imbalance at the oxidative level or control of the inflammatory response have been studied, to date it is not known exactly what molecular alteration is responsible for the appearance and development of FM. This situation makes extremely difficult the search for a curative treatment and a specific and accurate laboratory diagnosis. The concept of FIBROKIT was born with the intention of solving both shortcomings. Thus, the main objective of this project is to fully design the panel of specific biomarkers to use in FIBROKIT. To achieve this great objective, three specific objectives have been set:

1. Design, based on new scientific evidence, the components of the molecular diagnostic panel focused on Peripheral Blood Mononuclear Cell-specific proteins, plasma proteome and intestinal microbiome biomarkers.
2. Increase the number of patients participating in the study to validate the diagnostic capacity of the kit with robust statistics.
3. Analytically validate the response of the new panel of molecular markers to the course of the pathology and its monitoring under a nutritional intervention with olive oil-supplemented mediterranean diet.

• Study Type: Interventional
• Study Phase: Not Applicable

Study Design

Allocation: Randomized
Intervention Model: Parallel Assignment
Intervention Model Description:

Randomized, Double-blinded, placebo controlled intervention

Masking: Triple (Participant, Investigator, Outcomes Assessor)
Masking Description:

Treatment (extra virgin olive oil) and placebo (olive oil) were packed in 50 ml opaque envelopes for patients masking. Letters A and B were assigned randomly to each treatment by an external person not participating in the study design or analysis for investigator masking.

Primary Purpose: Diagnostic

• Condition: Fibromyalgia

Intervention

• Dietary Supplement: Extra Virgin Olive oil-supplemented Mediterranean Diet Intervention
  This intervention (treatment) consist of taking Extra Virgin Olive Oil ("treatment" group made up of 103 patients and 22 healthy people) for 6 months, accompanied by Mediterranean diet-based menus. During this time, blood and stool samples were taken at different points: before the start of the intervention (T0), 3 months after the start (T1), at the end of the intervention in the sixth month (T2) and 6 months after leaving the study (TF). Hence, a total of 4 time points were taken for analysis: T0, T1, T2, TF. Additionally, and following these same time points, the physical and psychological health status of the participants will be evaluated using the SF-36 health questionnaire. Similarly, the course of the pathology in terms of the Fibromyalgia Impact Questionnaire (FIQ) score were recorded.
• Dietary Supplement: Olive oil-supplemented Mediterranean Diet Intervention
  This intervention (treatment) consist of taking Olive Oil ("placebo" group made up of 103 patients and 22 healthy people) for 6 months, accompanied by Mediterranean diet-based menus. During this time, blood and stool samples were taken at different points: before the start of the intervention (T0), 3 months after the start (T1), at the end of the intervention in the sixth month (T2) and 6 months after leaving the study (TF). Hence, a total of 4 time points were taken for analysis: T0, T1, T2, TF. Additionally, and following these same time points, the physical and psychological health status of the participants will be evaluated using the SF-36 health questionnaire. Similarly, the course of the pathology in terms of the Fibromyalgia Impact Questionnaire (FIQ) score were recorded.

Study Arms

• Experimental: MED DIET + AOVE
  A six-months nutritional intervention with a Mediterranean diet, daily supplemented with 50 ml of extra virgin olive oil. The intake of the oil was divided in the main meals and needed to be consumed without cooking.
  Intervention: Dietary Supplement: Extra Virgin Olive oil-supplemented Mediterranean Diet Intervention
• Placebo Comparator: MED DIET + AO
  A six-months nutritional intervention with a Mediterranean diet, daily supplemented with 50 ml of olive oil. The intake of the oil was divided in the main meals and needed to be consumed without cooking.
  Intervention: Dietary Supplement: Olive oil-supplemented Mediterranean Diet Intervention

Publications *

• Martinez-Lara A, Moreno-Fernandez AM, Jimenez-Guerrero M, Diaz-Lopez C, De-Miguel M, Cotan D, Sanchez-Alcazar JA. Mitochondrial Imbalance as a New Approach to the Study of Fibromyalgia. Open Access Rheumatol. 2020 Aug 24;12:175-185. doi: 10.2147/OARRR.S257470. eCollection 2020.
• Ramirez-Tejero JA, Martinez-Lara E, Rus A, Camacho MV, Del Moral ML, Siles E. Insight into the biological pathways underlying fibromyalgia by a proteomic approach. J Proteomics. 2018 Aug 30;186:47-55. doi: 10.1016/j.jprot.2018.07.009. Epub 2018 Jul 17.
• Minerbi A, Fitzcharles MA. Gut microbiome: pertinence in fibromyalgia. Clin Exp Rheumatol. 2020 Jan-Feb;38 Suppl 123(1):99-104. Epub 2020 Feb 12.
• Clos-Garcia M, Andres-Marin N, Fernandez-Eulate G, Abecia L, Lavin JL, van Liempd S, Cabrera D, Royo F, Valero A, Errazquin N, Vega MCG, Govillard L, Tackett MR, Tejada G, Gonzalez E, Anguita J, Bujanda L, Orcasitas AMC, Aransay AM, Maiz O, Lopez de Munain A, Falcon-Perez JM. Gut microbiome and serum metabolome analyses identify molecular biomarkers and altered glutamate metabolism in fibromyalgia. EBioMedicine. 2019 Aug;46:499-511. doi: 10.1016/j.ebiom.2019.07.031. Epub 2019 Jul 18.

Recruitment Information

• Recruitment Status: Active, not recruiting
• Actual Enrollment (submitted: June 19, 2023): 250
• Original Actual Enrollment: Same as current
• Estimated Study Completion Date: December 31, 2023
• Estimated Primary Completion Date: August 31, 2023 (Final data collection date for primary outcome measure)

Eligibility Criteria

Inclusion Criteria:

• Women between 40 and 59 years
• Do not carry out any type of guided and structured physical activity (detailed in the exclusion criteria)
• Follow a balanced diet in terms of the food source (fruits, vegetables, legumes, meat, fish, etc.).

Exclusion Criteria:

• Being outside the established age range (40-59) at the time of the start of the study

• Have practiced some type of structured or planned physical activity more than 2 times a week during the last month, such as:

  1. Go to collective classes of Yoga, Tai-Chi, Zumba, dance, gymnastics or similar
  2. Going for a walk, cycling, hiking or similar for 30 minutes or more than 10,000 steps per day
• Being underweight (BMI < 18.5) or type II or higher obesity (BMI > 34.9)
• Suffer and have been diagnosed with any of the following chronic pathologies:

any type of cancer

• Acquired Immunodeficiency Syndrome (AIDS)
• Inflammatory diseases (rheumatoid arthritis, osteoarthritis)
• Gastrointestinal diseases (Crohn's disease, ulcerative colitis)
• Cardiovascular diseases (atherosclerosis, cardiomyopathy, stroke)
• Autoimmune diseases (systemic lupus erythematosus, celiac disease, Hashimoto's thyroiditis, multiple sclerosis)
• Metabolic diseases (Type I and II Diabetes, Metabolic Syndrome)
• Having been under intensive pharmacological treatment (3 or more drugs daily) with non-steroidal anti-inflammatory drugs, corticosteroids, analgesics, or antidepressants during the month prior to the start of the study.
• Being under antioxidant supplementation (Glutathione, Coenzyme Q10, plant extracts, phenolic compounds)
• Consuming an amount greater than 12 g/day of alcohol, admitted in the context of the Mediterranean diet (Willett et al., 1995).

Smoking or consuming any type of narcotic substance (regardless of the amount and frequency)

Sex/Gender

• Sexes Eligible for Study: Female
• Gender Based Eligibility: Yes

• Ages: 40 Years to 59 Years (Adult)
• Accepts Healthy Volunteers: Yes
• Contacts: Contact information is only displayed when the study is recruiting subjects
• Listed Location Countries: Spain

Administrative Information

• NCT Number: NCT05921409
• Other Study ID Numbers: IDI20210749; IDI-20210749 ( Other Grant/Funding Number: Centre for the Development of Industrial Technology, Ministry of Science and Innovation )
• Has Data Monitoring Committee: No

U.S. FDA-regulated Product

• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: No

IPD Sharing Statement

• Plan to Share IPD: No

• Current Responsible Party: Pronacera Therapeutics SL
• Original Responsible Party: Same as current
• Current Study Sponsor: Pronacera Therapeutics SL
• Original Study Sponsor: Same as current
• Collaborators: Centre for the Development of Industrial Technology, Spain

Investigators

• Study Chair: José Antonio Sánchez Alcázar, MD; Pablo de Olavide University, Seville (Spain)

• PRS Account: Pronacera Therapeutics SL
• Verification Date: July 2023