Autologous Serum-derived EV for Venous Trophic Lesions Not Responsive to Conventional Treatments (SER-VES-HEAL)

Autologous Serum-derived Extracellular Vesicles to Treat Venous Trophic Lesions Not Responsive to Conventional Treatments

Venous ulcers are defined by the presence of open lesions which represent the final stage of chronic venous disease or post-thrombotic syndrome. The risk factors for the development of venous ulcers include age, obesity, female sex, trauma, immobility, factor V mutation, thrombosis, venous agenesis.

Recommendation by the current guidelines includes compression and advanced dressing. However, in several cases, they fail to change patients' outcome. The aim of this study is to identify an alternative therapy to treat venous trophic lesions not responding to traditional therapeutic approaches using extracellular vesicles obtained from autologous serum.

Study Overview

• Status: Recruiting
• Conditions: Ulcer Venous
• Intervention / Treatment: Other: Autologous extracellular vesicles from serum

Detailed Description

In recent years, several in vitro and in vivo studies have provided been the beneficial effects of stem cells for tissue regeneration. Moreover, several randomized trials are ongoing and the interim results seem to give promising results and indicate their safety. Alternative approaches have recently been proposed, such as the injection of platelet preparation or the use of acellular amniotic membranes or extracellular vesicles (EV). EV act as cell-to-cell paracrine or endocrine-like communication mechanisms. The therapeutic potential of EV depends on the transfer of their cargo(proteins, RNAs, DNA, lipids, etc) to target cells (1). In the last decades, several pre-clinical studies have demonstrated safety. A pilot study was recently published on the use of EV as an effective treatment for retinal damage. At the University of Turin using a mouse model of ischemia/ reperfusion of the lower limb it was demonstrated that the intramuscular administration of EV obtained from the serum of healthy donors was associated with a higher density of local capillaries and an increase in laser Doppler signal compared to controls (2). A simple in vitro functional potency assay was developed, which allowed predicting EV therapeutic efficacy in vivo. The best response was obtained by intravenous administration immediately after surgical ligation and intramuscular in the next two days (T1 and T2). Through the Laser Doppler study, distal perfusion was evaluated after 7 days, which showed a clear improvement and protection of muscle damage. A more recent study has shown that even subjects with high cardiovascular risk (diabetic, obese, diabetic/obese and with arterial disease of the lower limbs) can benefit from the use of autologous EV (3).

The pilot study will involve the enrollment of 10 patients with bilateral venous ulcer refractory to conventional treatment for more than 6 months, recruited at the Vascular Ulcer Clinic of University Vascular Surgery. At the time of recruitment, venous Eco Doppler will be performed to evaluate the superficial and deep venous and arterial system to exclude possible "mixed" etiologies. A blood sample will then recovered for EV isolation in collaboration with the Blood Bank of the City of Health and Science of Turin. The characteristics of the ulcer will be entered in the database: dimensions (photo), margins, bottom and a swab for microbiology will be performed. Patients whose swab will be positive will be excluded.

EV will be analysed for their biological activity using the potency assay and will be used only if found to be effective (potency test). Subsequently, peri-wound injection of EV will be performed in a sterile environment. Sterile gauze and an elastic-compression bandage will be applied. As an internal control, the contralateral ulcer will be treated with a standard dressing and an elastic compression bandage. The patients will be evaluated on an outpatient basis on day 3 and then weekly. The results of treated patients will be collected in a dedicated database.

• Study Type: Interventional
• Enrollment (Anticipated): 10
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Maria Felice Brizzi, MD; Phone Number: +390116706653; Email: mariafelice.brizzi@unito.it
• Study Contact Backup: Name: Giovanni Camussi, MD; Phone Number: +390116709588; Email: giovanni.camussi@unito.it

Study Locations

• Italy
  • Turin, Italy, 10126
    • Recruiting
    • AOU San Giovanni Battista
    • Contact: Maria Felice Brizzi, MD; Phone Number: +390116706653; Email: mariafelice.brizzi@unito.it
    • Contact: Giovanni Camussi, MD; Phone Number: +390116709588; Email: giovanni.camussi@unito.it
    • Principal Investigator: Lorenzo Gibello, MD
    • Sub-Investigator: Margherita Alba Carlotta Pomatto, PhD
    • Principal Investigator: Maria Felice Brizzi, MD
    • Sub-Investigator: Anna Testa, MD
    • Sub-Investigator: Sergio D'Antico, MD

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 85 years (Adult, Older Adult)
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria: Adults and conscious patients

-

Exclusion Criteria:

• :> 85 years, diabetics, autoimmune diseases, neoplastic, arterial disease of the lower limbs.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Non-Randomized
• Interventional Model: Parallel Assignment
• Masking: Single

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Treatment; In the treatment arm, the lesion will be treated with EV for 3 weeks once a week	Other: Autologous extracellular vesicles from serum; Peri-wound injection of the vesicles will be performed in a sterile environment. Sterile gauze and an elastic-compression bandage will be applied.
Sham Comparator: Internal control; The contralateral ulcer will be treated with a standard dressing and an elastic-compression bandage for 3 weeks.	Other: Autologous extracellular vesicles from serum; Peri-wound injection of the vesicles will be performed in a sterile environment. Sterile gauze and an elastic-compression bandage will be applied.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes in the ulcer area from baseline to eight weeks	Ulcer areas will be recorded at baseline and weekly using digital camera. A standard rule was positioned on the surface of the skin around the ulcers to standardize the images and to allow data analysis. The ImageJ analysis program will be used to calculate the ulcer area. A blinded investigator manually delimitated the ulcer area to calculate the area. The total area will be expressed in square centimeters.	8 weeks

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Pain change	All patients should record their subjective feelings, including pain, burning and secretions. The level of pain will be monitored during the treatment as patient's experienced pain (decrease, unchange and/or increase). The amount of secretion will be classified as follow: absent, small, moderate, or abundant.	8 week

Collaborators and Investigators

• Sponsor: University of Turin, Italy
• Investigators: Principal Investigator: Maria Felice Brizzi, MD, University of Turin, Italy; Study Chair: Giovanni Camussi, MD, University of Turin, Italy

Publications and helpful links

General Publications

• Kholia S, Ranghino A, Garnieri P, Lopatina T, Deregibus MC, Rispoli P, Brizzi MF, Camussi G. Extracellular vesicles as new players in angiogenesis. Vascul Pharmacol. 2016 Nov;86:64-70. doi: 10.1016/j.vph.2016.03.005. Epub 2016 Mar 22.
• Cavallari C, Ranghino A, Tapparo M, Cedrino M, Figliolini F, Grange C, Giannachi V, Garneri P, Deregibus MC, Collino F, Rispoli P, Camussi G, Brizzi MF. Serum-derived extracellular vesicles (EVs) impact on vascular remodeling and prevent muscle damage in acute hind limb ischemia. Sci Rep. 2017 Aug 15;7(1):8180. doi: 10.1038/s41598-017-08250-0.
• Cavallari C, Figliolini F, Tapparo M, Cedrino M, Trevisan A, Positello L, Rispoli P, Solini A, Migliaretti G, Camussi G, Brizzi MF. miR-130a and Tgfbeta Content in Extracellular Vesicles Derived from the Serum of Subjects at High Cardiovascular Risk Predicts their In-Vivo Angiogenic Potential. Sci Rep. 2020 Jan 20;10(1):706. doi: 10.1038/s41598-019-55783-7.

Study record dates

Study Major Dates

• Study Start (Actual): September 18, 2020
• Primary Completion (Anticipated): December 30, 2023
• Study Completion (Anticipated): December 31, 2024

Study Registration Dates

• First Submitted: November 23, 2020
• First Submitted That Met QC Criteria: December 2, 2020
• First Posted (Actual): December 3, 2020

Study Record Updates

• Last Update Posted (Actual): May 12, 2023
• Last Update Submitted That Met QC Criteria: May 11, 2023
• Last Verified: May 1, 2023

More Information

Terms related to this study

• Keywords: serum; vascular disease; ulcers; Autologous EV
• Additional Relevant MeSH Terms: Cardiovascular Diseases; Vascular Diseases; Skin Diseases; Leg Ulcer; Skin Ulcer; Varicose Veins; Varicose Ulcer
• Other Study ID Numbers: CS2/1095/0090491

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