Functional Mitochondrial Analysis PBMCs (FMAP)

Functional Mitochondrial Analysis of Peripheral Blood Mononuclear Cells (PBMCs) - a Pilot Study

The primary goal of this prospective, exploratory, longitudinal, single-centre, cohort study is to assess the stability of the mitochondrial flux in PBMCs over long-term cryopreservation.

Secondary goals of this study are:

• to identify changes in the mitochondrial respiratory flux in different metabolic states of cryopreserved PBMCs during long-term cryopreservation.
• to assess variability between mitochondrial respiration from PBMCs isolated from same volunteers at different times, seasons or from different arms.

Study Overview

• Status: Active, not recruiting
• Conditions: Mitochondrial Function
• Intervention / Treatment: Other: Venipuncture; Other: Questionnaire

Detailed Description

The analysis of mitochondrial function can also be referred to as a bioenergetic snapshot. Mitochondria are dynamic metabolic organelles that adapt to various physiological demands, reflecting an individual's lifestyle and exposure to environmental factors, medications, and toxins. Numerous studies have shown that mitochondrial respiration declines with age and correlates with many age-related diseases. This raises the question of how mitochondria influence cells in a clinical context.

For this purpose, 20 participants are recruited and comprehensively characterized in terms of their demographic information and clinical profiles. Additionally, physical examinations are conducted, and participants are surveyed about their lifestyles through questionnaires. Over a 12-month period, blood samples are collected at intervals of three months, resulting in a total of five study visits. For the analysis of mitochondrial oxygen consumption, peripheral blood mononuclear cells (PBMCs) are preferably used, as they provide a minimally invasive and easily accessible insight into mitochondrial function and overall metabolic status and are isolated from the collected blood samples.

To enable the application of mitochondrial diagnostics in research for early disease detection and therapeutic development, additional information is needed regarding the stability of mitochondrial respiration in cryopreserved PBMCs using high-resolution respirometry (HRR) with O2k technology. The goal of this study is to assess how the duration of cryopreservation affects mitochondrial bioenergetics compared to freshly isolated PBMCs.

The study also considers a variety of parameters that could potentially influence the stability of mitochondrial respiration. These factors include non-fasting blood collection, discrepancies between the right and left arm, and seasonal effects. To what extent the intraindividual variability in these parameters affects the mitochondrial respiration is yet to be fully understood.

Furthermore, the longitudinal study design allows the tracking of mitochondrial activity and stability over time, providing a better understanding of the central processes of cellular respiration.

Thus, the planned study promises to yield significant insights into mitochondrial respiration and cellular bioenergetics in a clinical context.

• Study Type: Observational
• Enrollment (Estimated): 20

Contacts and Locations

Study Locations

• Austria
  • Tyrol
    • Innsbruck, Tyrol, Austria, 6020
      • Medical University Innsbruck - Department of Neurology

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

The source population for this study comprises adults (older than 18 years of age) with an equal distribution of male and female.

Overall, 20 individuals will be included in the study (10 male, 10 female).

Description

Inclusion Criteria:

• Aged between 18-85
• Willingness and ability to consent

Exclusion Criteria:

• Regular (e.g. daily, weekly or similar) intake of medication or nutritional supplements except oral and spiral contraceptives
• Autoimmune diseases or immune alterations
• Diseases in the context of haematopoiesis, haemophilia, hematophobia
• Diagnosed mild or major neurocognitive disorder
• Depressive episodes in the last two years
• Chronic infectious diseases
• Neurostimulators or drug pump
• Involved in competitive sports (over the past two years)
• Pregnancy

Study Plan

How is the study designed?

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Stability of mitochondrial respiratory flux (O2 flux) in cryopreserved PBMCs	Mitochondrial respiratory flux is assessed by High Resolution Respirometry analysis	1 week and every 8 weeks after cryopreservation
Stability of O2 concentration in cryopreserved PBMCs	Mitochondrial respiratory flux is assessed by High Resolution Respirometry analysis	1 week and every 8 weeks after cryopreservation
Assessment of mitochondrial respiratory flux (O2 flux) in fresh PBMCs compared to cryopreserved PBMCs	Mitochondrial respiratory flux is assessed by High Resolution Respirometry analysis	Baseline visit, 3, 6, 9, 12 months visit
Assessment of O2 concentration in fresh PBMCs compared to cryopreserved PBMCs	Mitochondrial respiratory flux is assessed by High Resolution Respirometry analysis	Baseline visit, 3, 6, 9, 12 months visit

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Assessment of O2 flux in fresh and cryopreserved PBMCs in fasted vs non-fasted sampling conditions	Assessement of mitochondrial bioenergetic snapshot in fresh and cryopreserved PBMCs at two different collection time points (fasted and non-fasted)	6 months visit
Assessment of O2 flux in fresh and cryopreserved PBMCs at different seasonal collection time points	Assessement of mitochondrial bioenergetic snapshot in fresh and cryopreserved PBMCs in different seasons	Baseline visit, 3, 6, 9, 12 months visit
Assessment of O2 flux in fresh and cryopreserved PBMCs at different venipuncture sites	Assessement of mitochondrial bioenergetic snapshot in fresh and cryopreserved PBMCs collected from left and right arm	3 months visit
Assessment of blood count and differential blood count I	Analysis of complete blood count (e.g. erythrocyte concentration (mg/dL))	Baseline visit, 3, 6, 9, 12 months visit
Assessment of blood count and differential blood count II	Analysis of complete blood count (e.g. haemoglobin concentration (g/dL))	Baseline visit, 3, 6, 9, 12 months visit
Concentration of Creatinine and Urea	Creatinine (mg/dL), Urea (mg/dL)	Baseline visit, 3, 6, 9, 12 months visit
Concentration of Creatine Kinase	Creatine Kinase (U/L)	Baseline, 6, 12 months visit
Concentration of Glucose	Glucose (mg/dL, mmol/L)	Baseline visit, 3, 6, 9, 12 months visit
Concentration of HbA1c	HbA1c (%)	Baseline, 6, 12 months visit
Concentration of Sodium, Potassium, Chloride and Calcium	Sodium (mmol/L), potassium (mmol/L), chloride (mmol/L), calcium (mmol/L)	Baseline, 6, 12 months visit
Concentration of GOT and GPT	Glutamic-oxaloacetic transaminase (GOT, U/L), glutamic-pyruvic transaminase (GPT, U/L), gamma-glutamyl-transpeptidase (gamma-GT, U/L), lactate dehydrogenase (LDH, U/L)	Baseline, 6, 12 months visit
Concentration of triglyceride and cholesterol	Triglyceride (mmol/L), cholesterol (all, mmol/L )	Baseline, 6, 12 months visit
Concentration of LDL-cholesterol and HDL-cholesterol	LDL-cholesterol (mg/dL), HDL-cholesterol (mg/dL)	Baseline, 6, 12 months visit
Concentration of Lipoprotein a	Lipoprotein a (mg/dL)	Baseline visit
Assessment of Sedimentation rate	Sedimentation rate (mm/h)	Baseline, 12 months visit
Concentration of C-reactive protein	CRP sensitive (mg/L)	Baseline visit, 3, 6, 9, 12 months visit
Concentration of Interleukin-6	Interleukin-6 (pg/ml)	Baseline, 6, 12 months visit
Concentration of Thyroid-stimulating hormone	TSH (mU/mL)	Baseline, 12 months visit
Concentration of Iric acid	Uric acid (mg/dL)	Baseline, 12 months visit
Concentration of Ferritin	Ferritin (ng/mL, μg/L)	Baseline, 12 months visit

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Demographic data I	Assessment of demographic data such as current age, sex at birth, location of birth	Baseline visit
Demographic data II	Assessment of demographic data such as: height (cm)	Baseline visit
Demographic data III	Assessment of demographic data such as: weight (kg)	Baseline visit
Demographic data - Personal background and lifestyle I	Assessment of demographic data with focus on personal background and lifestyle: e.g.; ethnicity (anamnesis); marital/relationship status (anamnesis); number of children (anamnesis); highest level of education (anamnesis)	Baseline visit
Menstrual cycle duration (women only)	Assessment of menstrual cycle with focus on cycle duration (in days)	Baseline visit, 3, 6, 9, 12 months visit
Menstrual cycle length (women only)	Assessment of menstrual cycle with focus on the initiation of the last cycle (day)	Baseline visit, 3, 6, 9, 12 months visit
Demographic data - Personal background and lifestyle II	Assessment of demographic data with focus on personal background and lifestyle: smoking and smoking history, alcohol consumption and diet	Baseline visit, 3, 6, 9, 12 months visit
Demographic data - Personal background and lifestyle III	Assessment of demographic data with focus on personal background and lifestyle: amount and intensity of physical activity	Baseline visit, 3, 6, 9, 12 months visit
Demographic data - Personal background and lifestyle IV	Assessment of personal background and lifestyle: sleep quality	Baseline visit, 3, 6, 9, 12 months visit
Medical History Assessment	Assessment of medical history: pre-existing illnesses; current illnesses or allergies; chronic illnesses; medication; previous surgical procedures within the last 2 years	Baseline visit, 3, 6, 9, 12 months visit
Assessment of mental well-being using the Hospital Anxiety and Depression Scale - German Version (HADS-D) questionnaire	The questionnaire uses an anxiety scale and a depression scale (0-21, each). The higher the value, the more anxious/depressed the patient. Cut-off for clinical significance: ≥8	Baseline visit, 3, 6, 9, 12 months visit

Collaborators and Investigators

• Sponsor: VASCage GmbH
• Collaborators: Oroboros Instruments
• Investigators: Principal Investigator: Michael Knoflach, AssozProf Dr, Medical University of Innsbruck

Study record dates

Study Major Dates

• Study Start (Actual): January 21, 2025
• Primary Completion (Estimated): January 1, 2026
• Study Completion (Estimated): May 1, 2026

Study Registration Dates

• First Submitted: June 18, 2024
• First Submitted That Met QC Criteria: July 4, 2024
• First Posted (Actual): July 12, 2024

Study Record Updates

• Last Update Posted (Actual): March 25, 2025
• Last Update Submitted That Met QC Criteria: March 3, 2025
• Last Verified: March 1, 2025

More Information

Terms related to this study

• Keywords: Mitochondrial respiration; PBMC
• Other Study ID Numbers: C_2_A.7-0039_009

Drug and device information, study documents

• 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