Efficacy of Intravenous Infusions of Stem Cells in the Treatment of COVID-19 Patients

Study Overview
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Brief Summary
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Stem cell therapy has emerged as a revolutionary treatment for diseases that were considered untreatable only a few years ago. Umbilical cord-derived mesenchymal stem cells (UCMSCs) have been shown to repair damaged liver, kidney, heart, pancreas, skin, cartilage, and cornea in animal models and several human trials. In addition to cellular replacement through regeneration, UCMSCs mediate through paracrine signaling pathways resulting in immune modulation. Clinical manifestations of coronavirus disease 2019 (COVID-19), are believed to arise from septic shock and cytokine storm that cause acute respiratory dysfunction and acute cardiac injury. There is presently no cure for the COVID-19 viral disease; however, multi-treatment strategies are being examined. During the past two months, four reports were published that suggest, mesenchymal stem cells (MSCs), owing to their powerful immunomodulatory ability, may prevent the cytokine storm and thus reduce the COVID-19 related morbidity. All studies reported that COVID-19 patients responded favorably to MSCs therapy. These reports, taken together with the previous successes of stem cell therapy in animal models, the investigators, a seven-institution consortium, propose to explore the efficacy of UCMSC treatment in COVID-19 patients at Jinnah hospital, Lahore. The investigators propose to administer UCMSCs in patients with acute pulmonary inflammation due to COVID-19 infection with moderate to severe symptoms. In the first cohort of 15 patients, UCMSCs will be administered with three intravenous infusions of 500,000 UCMSCs per Kg body weight each on days 1, 3, and 5. The second group of five patients serving as control will only receive standard treatment. During the 30-day post-infusion period, a battery of tests will be performed to evaluate the safety and efficacy of the UCMSCs treatment. In parallel, the investigators propose a comparative study to determine COVID-19 viral count by quantitative real-time PCR and through viral coat protein ELISA, developed in the investigator advisor lab (Dr. Tauseef Butt, Progenra Inc. Philadelphia, USA) with the ultimate objective to locally developing a rapid diagnostic assay.

Detailed Description
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Isolation and characterization of human umbilical cord-derived mesenchymal stem cells (UCMSCs): Human umbilical cord tissues along with informed consent forms will be collected from hepatitis B, C, and COVID-19 virus-negative women with healthy pregnancies during the Cesarean Section surgery after completion of gestation period. The cord tissue will be transported in sterile 1x phosphate-buffered saline (PBS) containing 200 units/ml penicillin and 200 µg/ml streptomycin on ice. In the biosafety cabinet, the cord will be washed with 4-5 changes of sterile 1x PBS and placed in a Petri plate with 15 ml PBS. The cord will be gently scraped with a surgical blade to remove any dead cells. A 9 cm umbilical cord will be cut into three equal pieces and wash thoroughly to remove blood clots, umbilical cord arteries, and veins. Segments will be washed three times with PBS and minced. The minced pieces will be incubated in 17.5 ml of collagenase solution (201 U/ml collagenase type I in serum-free DMEM-HG) in a 50 ml conical tube for  3.5 hours in an incubator at 5% CO2, 95% humidity at 37⁰C. After  3.5 hours, the digested lysate will pass through a strainer and will be diluted three times with 1x PBS. Following centrifugation, the cells will be seeded into two 25 cm2 flasks and will be placed in an incubator at 5% CO2, 95% humidity at 37⁰C. The flasks will be fed with fresh media (DMEM-HG supplemented with 20% FBS and 1% antibiotic solution) every third day. At around day 18, cells will reach up to 85% confluency and will be transferred in two 75 cm2 flasks with media replaced at alternate days. UCMSCs at P3 will be characterized using different specific antibodies. Cells at P3 will be used for transplantation in patients. Fifteen (15) subjects will be treated with three intravenous infusions (IV) of 500,000 UCMSCs per Kg body weight delivered via peripheral intravenous infusion.

Official Title
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Efficacy of Intravenous Infusions of Stem Cells in the Treatment of COVID-19 Patients

Conditions
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Corona Virus Infection

Intervention / Treatment
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* Biological: Intravenous Infusions of Stem Cells


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Provide written informed consent Male or female subjects age >18 years at the time of signing the Informed Consent Form Must have a clinical diagnosis of COVID-19, with at least one of clinical symptoms (e.g., fever ≥38°C, fatigue, cough) and a positive result by the reverse-transcription polymerase chain reaction (RT-PCR) testing or equivalent Individuals with moderate to severe COVID-19 symptoms Adequate venous access For female patients only, willingness to use recommended birth control until 6 months post-treatment Must agree to comply with all study requirements and be willing to complete all study visits Need in-patient admission Exclusion Criteria: A female who is pregnant, nursing, or of child-bearing potential while not practicing effective contraceptive methods. Female subjects must undergo a blood pregnancy test at screening and prior to infusion. Inability to perform any of the assessments required for endpoint analysis Have known allergies to penicillin or streptomycin Have a clinical history of malignancy within 3 years (i.e., subjects with prior malignancy must be disease-free for 3 years), except curatively-treated basal cell carcinoma, squamous cell carcinoma, melanoma in situ or cervical carcinoma, if recurrence occurs; - History of drug abuse (illegal street drugs except for marijuana, or prescription medications not being used appropriately for a pre-existing medical condition) or alcohol abuse (≥ 5 drinks/day for ˃ 3 months), or documented medical, occupational, or legal problems arising from the use of alcohol or drugs within the past 24 months Be serum positive for HIV, hepatitis BsAg or Viremic hepatitis C.

Ages Eligible for Study
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Minimum Age: 30 Years
Maximum Age: 70 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: Group 1 (15 subjects) Fifteen (15) subjects will be treated with three intravenous infusion (IV) of 5 x 10^5 (per Kg body weight) UCMSCs delivered via peripheral intravenous infusion on days 1, 3 and 5 besides the standard care (SOC). Group 2 (5 subjects) Five (5) subjects will be treated under SOC. From two subjects in group 1 will not be treated 24hrs apart. Patients will be randomized in a 1:3 ratio (SOC: UCMSC).
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Group 1 : Treatment<br>Fifteen (15) subjects will be treated with three intravenous infusion (IV) of 5 x 10^5 UCMSCs per Kg body weight delivered via peripheral intravenous infusion on days 1, 3 and 5 besides the standard care (SOC). | Biological: Intravenous Infusions of Stem Cells<br>* Cultured stem cells will be injected at days 1, 3 and 5 intravenously. While patients will be monitored and evaluated for various hematological and organ functions at baseline (day 0) and days 1, 3, 5, 7, 14, 21 and 30<br>|
| No Intervention: Group 2: standard care<br>Five (5) subjects will be treated under Standard of Care (SOC) . | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Safety and efficacy assessment of infusion associated adverse events | Number of participants with significant side effects in stem cell treated group | Day 01 to Day 30 | 
| Chest Radiograph or Chest CT Scan | Assessment of Pneumonia improvement as a result of stem cell infusions | Day 01 to Day 30 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| COVID-19 Quantitative Real Time PCR | Quantitative real-time PCR analysis for the evaluation of negative corona virus test results following stem cell treatment | Day 01 to Day 30 | 
| Sequential Organ Failure Assessment (SOFA) Score | Evaluation of organ function (Each organ system is assigned a value for 0 (normal) to 4 (highest degree of dysfunction)) | Day 01 to Day 30 | 
| Rate of mortality | Number of all mortalities within 30 days post first infusion | Day 01 to Day 30 | 
| Clinical Respiratory Changes | Examination of improvement in the physiology of lungs after treatment | Day 01 to Day 30 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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COVID-19, Stem Cells treatment