Mitigating Mitochondrial RNA Release During Aging to Control Inflammation and Senescence (MIRACLE)

Mitigating Mitochondrial RNA Release During Aging to Control Inflammation and Senescence, Preserving Organ Function and Enhancing Healthspan

The MIRACLE study aims to investigate age-related mitochondrial dysfunction, mitochondrial RNA (mtRNA) release, inflammation, and cellular senescence in adult participants across three age groups. Skin-derived fibroblasts and peripheral blood mononuclear cells (PBMCs) will be isolated from skin biopsy and blood samples to characterize age-related cellular and molecular changes and to test experimental therapeutic strategies identified in preclinical studies. Serum, plasma, and whole-blood RNA will be used for protocol-defined analyses of circulating inflammatory mediators and systemic transcriptional signatures related to inflammation, type I interferon activation, mitochondrial stress response, immune aging, and senescence-associated pathways.

Study Overview

• Status: Not yet recruiting
• Conditions: Inflammation; Aging; Healthy Ageing
• Intervention / Treatment: Procedure: Skin biopsy and venous blood sampling

Detailed Description

Mitochondria are crucial for ATP production and intracellular signaling in higher eukaryotic cells. These organelles contain genetic material that reflects their bacterial ancestry, including mitochondrial RNA (mtRNA). Under normal conditions, mtRNA is tightly confined and processed within mitochondria, ensuring the proper synthesis of proteins required for oxidative phosphorylation. Recent evidence suggests that mitochondrial stress may promote the leakage of mitochondrial components, including mtRNA. Once released into the cytosol, mtRNA can be sensed by cytosolic pattern recognition receptors (PRRs), thereby activating signaling pathways that promote the production of pro-inflammatory cytokines.

Although these mechanisms have been investigated mainly under conditions of acute stress, the specific contribution of mitochondrial dysfunction and mtRNA release to chronic low-grade inflammation and cellular senescence during physiological aging remains incompletely understood. This represents an important knowledge gap in the understanding of molecular processes that may contribute to age-related inflammation and functional decline.

The MIRACLE project includes a broade series of preclinical in vitro and in vivo experiments aimed at clarifying the mechanisms linking mitochondrial dysfunction, mtRNA release, inflammatory pathway activation, and cellular senescence during aging. These experimental activities are designed to define the biological pathways involved and to identify potential strategies capable of modulating mtRNA-associated inflammatory and senescence responses.

Within this broader framework, the present human study is intended to corroborate and extend the preclinical findings in humans. To this aim, adult participants across different age groups will be enrolled, and skin biopsy and blood samples will be collected to obtain skin-derived fibroblasts, peripheral blood mononuclear cells (PBMCs), serum, plasma, and whole-blood RNA.

Skin-derived fibroblasts will provide an accessible primary cell model for the investigation of age-related cellular and molecular changes. Fibroblasts isolated from participants of different ages will be used to assess mitochondrial function, mtRNA release, inflammatory signaling, and markers of cellular senescence. PBMCs collected from the same participants will be analyzed as a complementary blood-derived cellular model to evaluate systemic immune and inflammatory features, including immunosenescence-related signatures. Serum and plasma samples will be used to measure circulating inflammatory mediators and senescence-associated factors, while whole-blood RNA will be used to assess systemic transcriptional signatures related to inflammation, type I interferon activation, mitochondrial stress response, immune aging, and senescence-associated pathways.

• Study Type: Interventional
• Enrollment (Estimated): 90
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Luca Perico, PhD; Phone Number: +3903542131; Email: luca.perico@marionegri.it
• Study Contact Backup: Name: Miriam Rigoldi, MD; Phone Number: +390354535327; Email: miriam.rigoldi@marionegri.it

Study Locations

• Italy
  • BG
    • Bergamo, BG, Italy, 24126
      • Laboratory for Targeted Therapy in Autoimmune Diseases
      • Contact: Luca Perico, PhD; Phone Number: +3003542131; Email: luca.perico@marionegri.it
    • Ranica, BG, Italy, 24020
      • Centro di Ricerche Cliniche per le Malattie Rare "Aldo e Cele Daccò"
      • Contact: Miriam Rigoldi, MD; Phone Number: +390354535327; Email: miriam.rigoldi@marionegri.it

Participation Criteria

Eligibility Criteria

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

Description

Inclusion Criteria:

• Male and female
• Age between 18 and 90 years (stratified in three groups: young, middle-aged, elderly)
• Written informed consent

Exclusion Criteria:

• Inability to understand the potential risk and benefits of the study
• Legal incapacity
• Subjects who have taken antibiotics, anti-inflammatory drugs, or antihistamines within the past 7 days
• Diagnosis of diabetes mellitus
• Use of anticoagulant medications
• Any subject with a contraindication to the mini-invasive biopsy procedure

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Non-Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Young adults; Female and male volunteers aged 18-40 years	Procedure: Skin biopsy and venous blood sampling; A single 3-4 mm punch biopsy of forearm skin is performed under local anesthesia (lidocaine 1% with epinephrine 1:100,000) for primary dermal fibroblast isolation. Venous blood is collected by standard phlebotomy from the antecubital fossa (43 mL total volume per participant): 5 mL in SST tube for serum separation, 3 mL in EDTA tube for plasma separation, 32 mL in EDTA tubes for PBMC isolation by density gradient centrifugation, and 3 mL in Tempus Blood RNA tube for whole-blood RNA stabilization. All procedures are performed at a single visit at enrollment. Samples are processed for biospecimen-based ex vivo molecular and cellular analyses; no investigational product is administered to participants.
Experimental: Middle-aged adults; Female and male volunteers aged 40-60 years	Procedure: Skin biopsy and venous blood sampling; A single 3-4 mm punch biopsy of forearm skin is performed under local anesthesia (lidocaine 1% with epinephrine 1:100,000) for primary dermal fibroblast isolation. Venous blood is collected by standard phlebotomy from the antecubital fossa (43 mL total volume per participant): 5 mL in SST tube for serum separation, 3 mL in EDTA tube for plasma separation, 32 mL in EDTA tubes for PBMC isolation by density gradient centrifugation, and 3 mL in Tempus Blood RNA tube for whole-blood RNA stabilization. All procedures are performed at a single visit at enrollment. Samples are processed for biospecimen-based ex vivo molecular and cellular analyses; no investigational product is administered to participants.
Experimental: Elderly adults; Female and male volunteers aged 60-90 years	Procedure: Skin biopsy and venous blood sampling; A single 3-4 mm punch biopsy of forearm skin is performed under local anesthesia (lidocaine 1% with epinephrine 1:100,000) for primary dermal fibroblast isolation. Venous blood is collected by standard phlebotomy from the antecubital fossa (43 mL total volume per participant): 5 mL in SST tube for serum separation, 3 mL in EDTA tube for plasma separation, 32 mL in EDTA tubes for PBMC isolation by density gradient centrifugation, and 3 mL in Tempus Blood RNA tube for whole-blood RNA stabilization. All procedures are performed at a single visit at enrollment. Samples are processed for biospecimen-based ex vivo molecular and cellular analyses; no investigational product is administered to participants.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Age-dependent differences in cytosolic release of mitochondrial RNA (mtRNA) in primary human fibroblasts	Assessment of release of mtRNA in the cytosol of primary fibroblasts isolated from study participants across age groups. Cytosolic mtRNA levels assessed through analysis of the mean fluorescence intensity (MFI) per cell by immunofluorescence.	Day 1 (at enrollment)
Age-dependent differences in mitochondrial function in primary human fibroblasts	Characterization of mitochondrial function parameters in primary fibroblasts isolated from study participants across age groups. Mitochondrial function assessed through analysis of mitochondrial Complex I activity in nmol/min/mg protein by functional ELISA.	Day 1 (at enrollment)
Activation of inflammatory pathways in primary human fibroblasts	Assessment of inflammatory pathway activation markers in fibroblasts isolated from participants across age groups. Inflammatory pathway activation assessed through Interleukin-6 concentration in skin-derived fibroblast culture supernatant in pg/mL	Day 1 (at enrollment)
Activation of senescence pathways in primary human fibroblasts	Assessment of cellular senescence markers and senescence-associated pathway activation in fibroblasts isolated from participants across age groups. Senescence pathway activation assessed as percentage (%) of Senescence-associated β-galactosidase (SA-β-gal)-positive cells per microscopic field.	Day 1 (at enrollment)
Comparison of fibroblast findings with PBMC-derived parameters from the same participants	Integration and comparison of fibroblast-derived molecular data with corresponding PBMC-derived measurements obtained from the same participants as described above.	Day 1 (at enrollment)

Collaborators and Investigators

• Sponsor: Mario Negri Institute for Pharmacological Research
• Investigators: Study Director: Giuseppe Remuzzi, Istituto Di Ricerche Farmacologiche Mario Negri

Publications and helpful links

General Publications

• Zuber TJ. Punch biopsy of the skin. Am Fam Physician. 2002 Mar 15;65(6):1155-8, 1161-2, 1164.
• Nischal U, Nischal Kc, Khopkar U. Techniques of skin biopsy and practical considerations. J Cutan Aesthet Surg. 2008 Jul;1(2):107-11. doi: 10.4103/0974-2077.44174.
• Yasui Y, Kato H, Oda T, Nakamura M, Morita A. Complications and risk factors of punch biopsy: A retrospective large-scale study. J Dermatol. 2023 Jan;50(1):98-101. doi: 10.1111/1346-8138.16585. Epub 2022 Sep 24.
• Akiyama Y, Norimatsu Y, Ohno Y. Prophylactic antimicrobials may not be needed to prevent surgical site infection after skin biopsy: a retrospective study. Antimicrob Resist Infect Control. 2022 Feb 16;11(1):35. doi: 10.1186/s13756-022-01077-z.

Study record dates

Study Major Dates

• Study Start (Estimated): September 1, 2026
• Primary Completion (Estimated): September 1, 2031
• Study Completion (Estimated): September 1, 2031

Study Registration Dates

• First Submitted: April 29, 2026
• First Submitted That Met QC Criteria: May 12, 2026
• First Posted (Actual): May 19, 2026

Study Record Updates

• Last Update Posted (Actual): May 19, 2026
• Last Update Submitted That Met QC Criteria: May 12, 2026
• Last Verified: May 1, 2026

More Information

Terms related to this study

• Keywords: mitochondrial dysfunction; PBMCs; inflammaging; senescence; skin-derived fibroblasts; mtRNA
• Additional Relevant MeSH Terms: Pathologic Processes; Metabolic Diseases; Pathological Conditions, Signs and Symptoms; Nutritional and Metabolic Diseases; Inflammation; Mitochondrial Diseases; Investigative Techniques; Specimen Handling; Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Punctures; Surgical Procedures, Operative; Blood Specimen Collection
• Other Study ID Numbers: MIRACLE; FIS-2024-00858 (Other Grant/Funding Number: Italian Ministry of University and Research)

Plan for Individual participant data (IPD)

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

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No