CALM-AF-AI: Counteracting Age-related Loss of Muscle With AAV-Follistatin Combined With Angiogenesis-Inducing VEGF Plasmid Gene Therapy

NCT07443826 | Recruiting Phase 1

• 1 other identifier: UBL-CALMAF-001
• Study Type: interventional
• Target: 12
• Locations: 1 country
• Sites: 1

Brief Summary

This Phase 1/2a, open-label, non-randomized study is designed to evaluate the safety and tolerability of intramuscular AAV9-Follistatin gene therapy administered either as monotherapy or in combination with a VEGF-encoding plasmid. Secondary objectives include the assessment of preliminary signals of biological and functional activity, including changes in skeletal muscle mass and performance.

Conditions

Age-related Muscle Decline

Keywords

Sarcopenia; Muscle Atrophy; Skeletal Muscle; Muscle Weakness; Muscle Strength; Aging; Frailty; Follistatin; Myostatin; Viral Vectors; Adeno-Associated Viruses; Activins; Vascular Endothelial Growth Factor A; Angiogenesis Inducing Agents; Plasmids; Intramuscular Injections; Immunosuppression; Prednisolone; Sirolimus; Safety; Dual-Energy X-Ray Absorptiometry; Physical Endurance; Quality of Life; Adults; Middle Aged; Gene Therapy; Grip strength; Rapamycin; 6MWT; VO2max; 1RM

Outcome Measures

Primary Outcomes (2)

• Number of participants with treatment-emergent adverse events (TEAEs), including serious adverse events (SAEs)

  Incidence, severity, and relatedness of treatment-emergent adverse events (TEAEs), including serious adverse events (SAEs), through Day 90, graded according to CTCAE

  Day 1 through Day 90 after AAV administration

• Number of participants with dose-limiting toxicities (DLTs)

  Incidence of protocol-defined dose-limiting toxicities (DLTs) within the 21-day DLT observation window following AAV administration

  Day 1 through Day 21 after AAV administration

Secondary Outcomes (4)

• Number of participants with predefined clinically significant laboratory abnormalities

  Day 1 through Day 90 after AAV administration

• Number of participants with new-onset symptomatic heart failure or arrhythmias (CTCAE Grade ≥2)

  Day 1 through Day 90 after AAV administration

• Number of participants with injection site reactions

  Day 1 through Day 90 after AAV administration

• Number of participants who discontinue study treatment due to adverse events

  Day 1 through Day 90 after AAV administration

Other Outcomes (14)

• Change from baseline in Appendicular Lean Mass Index (ALMI) measured by DXA

  Baseline through Month 12

• Change from baseline in Bone Mineral Density (BMD) measured by DXA (g/cm²)

  Baseline through Month 12

• Change from baseline in lower limb one-repetition maximum (1RM) strength (kg)

  Baseline through Month 12

Study Arms (3)

Low-Dose AAV-Follistatin Monotherapy

EXPERIMENTAL

Participants will receive a single intramuscular dose of AAV-Follistatin (5 × 10¹⁰ vg/kg) on Day 1. The total dose will be administered bilaterally across predefined injection sites in major limb muscles (quadriceps femoris, gluteus maximus, gastrocnemius, and biceps brachii) according to a standardized injection map. Rapamycin will be given per protocol for approximately two months to mitigate immune responses to the AAV vector. This sentinel cohort follows a sequential 3+3 dose-escalation design; dose-limiting toxicities will be assessed through Day 21 prior to escalation to Arm 2.

Genetic: AAV9-Follistatin gene therapy

High-Dose AAV-Follistatin Monotherapy

EXPERIMENTAL

After safety review of Arm 1, participants in Arm 2 will receive a single higher intramuscular dose of AAV-Follistatin (1 × 10¹¹ vg/kg) on Day 1, administered bilaterally across predefined injection sites in major limb muscles according to the same standardized injection map. Prophylactic immunomodulation with rapamycin will be provided per protocol for approximately two months, with identical safety monitoring procedures as in Arm 1. This cohort is intended to evaluate dose-dependent safety and tolerability of AAV-Follistatin monotherapy. Dose-limiting toxicities (DLTs) will be assessed through Day 21, and escalation to Arm 3 will proceed following review of safety data and confirmation of predefined criteria.

Genetic: AAV9-Follistatin gene therapy

Combination: AAV-Follistatin + VEGF Plasmid

EXPERIMENTAL

Following safety review of Arm 2, participants in Arm 3 will receive VEGF plasmid (total dose 4.8 mg) administered intramuscularly on Day 1 and Day 12 (±2 days) across the same predefined bilateral muscle groups used for AAV-Follistatin (quadriceps femoris, gluteus maximus, gastrocnemius, and biceps brachii). AAV-Follistatin will then be administered 15 ± 1 days after the second VEGF dose (approximately Day 27-29) using the identical standardized injection map. The AAV dose for Arm 3 will be selected based on safety review of prior cohorts and will be either 5 × 10¹⁰ vg/kg or 1 × 10¹¹ vg/kg. Rapamycin will be provided per protocol for approximately two months beginning around the time of AAV administration to mitigate potential immune responses. This cohort evaluates the safety and feasibility of sequential VEGF-mediated vascular support combined with myoanabolic AAV-Follistatin gene therapy.

Genetic: AAV9-Follistatin gene therapy; Genetic: VEGF Plasmid

Interventions

AAV9-Follistatin gene therapy GENETIC

One-time intramuscular administration of an adeno-associated virus, serotype 9, (AAV9) vector encoding human follistatin.

Combination: AAV-Follistatin + VEGF Plasmid; High-Dose AAV-Follistatin Monotherapy; Low-Dose AAV-Follistatin Monotherapy

VEGF Plasmid GENETIC

Intramuscular supercoiled plasmid DNA gene therapy encoding vascular endothelial growth factor (VEGF).

Combination: AAV-Follistatin + VEGF Plasmid

Eligibility Criteria

• Age: 35 Years - 75 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• Voluntary written informed consent obtained prior to any study-related procedures
• Ability to read, understand, and sign the Informed Consent Form and reliably complete required study documents
• Willingness to undergo medical intervention, including genetic therapy, and to comply with the visit schedule and all study procedures
• Commitment to maintain a stable medication and supplement regimen throughout the study, with no initiation of new medications, supplements, or performance-enhancing substances unless approved by the Investigator
• Men and women aged 35-75 years
• Body mass index (BMI) between 18.0 and 35.0 kg/m² at screening
• Active Prospera ZEDE eResidency or Physical Residency
• Stable comorbid conditions for at least 3 months prior to screening
• Postmenopausal status (women)
• Willingness to use reliable contraception for 6 months following therapy
• Low or undetectable antibody titers to AAV9 (≤1:100 by ELISA)

You may not qualify if:

• Pregnancy, breastfeeding, or intent to become pregnant; premenopausal status (unless ≥12 months amenorrhea or FSH ≥30 IU/L)
• Subjects who have a history of alcohol or drug abuse within 1 year of study entry
• Initiation of prohibited medications, supplements, or interventions during the study period that may confound efficacy or safety assessments
• Active malignancy or ANY history of cancer
• Strong family history of cancer in first-degree relatives (≥2 relatives with cancer diagnosed \<60 years) OR known hereditary cancer syndrome (BRCA1/2, Lynch syndrome, Li-Fraumeni, FAP, HNPCC) without genetic counseling and enhanced surveillance clearance
• Clinically significant cardiovascular disease, including:
• Any history of stroke, transient ischemic attack (TIA), myocardial infarction (MI), or unstable angina (regardless of time since event)
• Diagnosed coronary artery disease (CAD) documented by coronary angiography or stress testing
• Significant atherosclerotic disease at any location (stenosis ≥50% in carotid, femoral, or other major arteries)
• Prior coronary revascularization (percutaneous coronary intervention \[PCI\] or coronary artery bypass grafting \[CABG\])
• Prior valvular repair or replacement
• History or current diagnosis of heart failure
• Uncontrolled hypertension (SBP \>140 mmHg or DBP \>85 mmHg despite treatment)
• Left ventricular ejection fraction (LVEF) \<50%, QTc ≥480 ms, or severe valvular heart disease
• Ventricular arrhythmias requiring chronic drug treatment or implantable cardioverter-defibrillator

Sponsors & Collaborators

• Unlimited Biotechnology LLC: lead
• Global Alliance for Regenerative Medicine: collaborator

Study Sites (1)

GARM

Coxen Hole, Bay Islands, 34101, Honduras

RECRUITING

Related Publications (11)

• Suoranta T, Laham-Karam N, Yla-Herttuala S. Strategies to improve safety profile of AAV vectors. Front Mol Med. 2022 Nov 1;2:1054069. doi: 10.3389/fmmed.2022.1054069. eCollection 2022.

  PMID: 39086961 BACKGROUND

• Tang R, Harasymowicz NS, Wu CL, Collins KH, Choi YR, Oswald SJ, Guilak F. Gene therapy for follistatin mitigates systemic metabolic inflammation and post-traumatic arthritis in high-fat diet-induced obesity. Sci Adv. 2020 May 8;6(19):eaaz7492. doi: 10.1126/sciadv.aaz7492. eCollection 2020 May.

  PMID: 32426485 BACKGROUND

• Mendell JR, Sahenk Z, Al-Zaidy S, Rodino-Klapac LR, Lowes LP, Alfano LN, Berry K, Miller N, Yalvac M, Dvorchik I, Moore-Clingenpeel M, Flanigan KM, Church K, Shontz K, Curry C, Lewis S, McColly M, Hogan MJ, Kaspar BK. Follistatin Gene Therapy for Sporadic Inclusion Body Myositis Improves Functional Outcomes. Mol Ther. 2017 Apr 5;25(4):870-879. doi: 10.1016/j.ymthe.2017.02.015. Epub 2017 Mar 6.

  PMID: 28279643 BACKGROUND

• Nance ME, Shi R, Hakim CH, Wasala NB, Yue Y, Pan X, Zhang T, Robinson CA, Duan SX, Yao G, Yang NN, Chen SJ, Wagner KR, Gersbach CA, Duan D. AAV9 Edits Muscle Stem Cells in Normal and Dystrophic Adult Mice. Mol Ther. 2019 Sep 4;27(9):1568-1585. doi: 10.1016/j.ymthe.2019.06.012. Epub 2019 Jul 3.

  PMID: 31327755 BACKGROUND

• Al-Zaidy SA, Sahenk Z, Rodino-Klapac LR, Kaspar B, Mendell JR. Follistatin Gene Therapy Improves Ambulation in Becker Muscular Dystrophy. J Neuromuscul Dis. 2015 Sep 2;2(3):185-192. doi: 10.3233/JND-150083.

  PMID: 27858738 BACKGROUND

• Deev R, Plaksa I, Bozo I, Isaev A. Results of an International Postmarketing Surveillance Study of pl-VEGF165 Safety and Efficacy in 210 Patients with Peripheral Arterial Disease. Am J Cardiovasc Drugs. 2017 Jun;17(3):235-242. doi: 10.1007/s40256-016-0210-3.

  PMID: 28050885 BACKGROUND

• Deev R, Plaksa I, Bozo I, Mzhavanadze N, Suchkov I, Chervyakov Y, Staroverov I, Kalinin R, Isaev A. Results of 5-year follow-up study in patients with peripheral artery disease treated with PL-VEGF165 for intermittent claudication. Ther Adv Cardiovasc Dis. 2018 Sep;12(9):237-246. doi: 10.1177/1753944718786926. Epub 2018 Jul 11.

  PMID: 29996720 BACKGROUND

• Kota J, Handy CR, Haidet AM, Montgomery CL, Eagle A, Rodino-Klapac LR, Tucker D, Shilling CJ, Therlfall WR, Walker CM, Weisbrode SE, Janssen PM, Clark KR, Sahenk Z, Mendell JR, Kaspar BK. Follistatin gene delivery enhances muscle growth and strength in nonhuman primates. Sci Transl Med. 2009 Nov 11;1(6):6ra15. doi: 10.1126/scitranslmed.3000112.

  PMID: 20368179 BACKGROUND

• Mendell JR, Sahenk Z, Malik V, Gomez AM, Flanigan KM, Lowes LP, Alfano LN, Berry K, Meadows E, Lewis S, Braun L, Shontz K, Rouhana M, Clark KR, Rosales XQ, Al-Zaidy S, Govoni A, Rodino-Klapac LR, Hogan MJ, Kaspar BK. A phase 1/2a follistatin gene therapy trial for becker muscular dystrophy. Mol Ther. 2015 Jan;23(1):192-201. doi: 10.1038/mt.2014.200. Epub 2014 Oct 17.

  PMID: 25322757 BACKGROUND

• Giesige CR, Wallace LM, Heller KN, Eidahl JO, Saad NY, Fowler AM, Pyne NK, Al-Kharsan M, Rashnonejad A, Chermahini GA, Domire JS, Mukweyi D, Garwick-Coppens SE, Guckes SM, McLaughlin KJ, Meyer K, Rodino-Klapac LR, Harper SQ. AAV-mediated follistatin gene therapy improves functional outcomes in the TIC-DUX4 mouse model of FSHD. JCI Insight. 2018 Nov 15;3(22):e123538. doi: 10.1172/jci.insight.123538.

  PMID: 30429376 BACKGROUND

• Iyer CC, Chugh D, Bobbili PJ, Iii AJB, Crum AE, Yi AF, Kaspar BK, Meyer KC, Burghes AHM, Arnold WD. Follistatin-induced muscle hypertrophy in aged mice improves neuromuscular junction innervation and function. Neurobiol Aging. 2021 Aug;104:32-41. doi: 10.1016/j.neurobiolaging.2021.03.005. Epub 2021 Mar 12.

  PMID: 33964607 BACKGROUND

MeSH Terms

Conditions

Sarcopenia; Muscular Atrophy; Muscle Weakness; Frailty

Condition Hierarchy (Ancestors)

Neuromuscular Manifestations; Neurologic Manifestations; Nervous System Diseases; Atrophy; Pathological Conditions, Anatomical; Pathological Conditions, Signs and Symptoms; Signs and Symptoms; Muscular Diseases; Musculoskeletal Diseases; Pathologic Processes

Central Study Contacts

Ivan Morgunov

CONTACT

+31623454906; ivan@unlimit.bio

Study Design

• Study Type: interventional
• Phase: phase 1
• Allocation: NON RANDOMIZED
• Masking: NONE
• Purpose: PREVENTION
• Intervention Model: SEQUENTIAL
• Sponsor Type: INDUSTRY
• Responsible Party: SPONSOR

Study Record Dates

• First Submitted: December 4, 2025
• First Posted: March 2, 2026
• Study Start: June 1, 2026
• Primary Completion (Estimated): September 30, 2027
• Study Completion (Estimated): June 30, 2028
• Last Updated: July 13, 2026

Record last verified: 2026-07

Locations

HO (1)