Phase I Evaluation of Autologous CD34+ Cells Transduced with Lentiviral Vector Encoding Codon-Optimized PKLR Gene in Pyruvate Kinase Deficiency
- Trial ID
- 2024-511520-13-00
- Protocol
- RP-L301-0119
- Sponsor
- Rocket Pharmaceuticals Inc.
Trial statistics
Diseases & Conditions
Objectives
The primary objective of this Phase I clinical trial is to evaluate the **safety** and toxicity associated with the infusion of autologous CD34+ cells transduced with a lentiviral vector carrying the codon-optimized red cell pyruvate kinase (coRPK) gene. This investigational product is intended for use in adult and pediatric subjects with **Pyruvate Kinase Deficiency** (PKD), an inherited metabolic disorder affecting the survival of red blood cells. The clinical relevance of this objective lies in its potential to provide a novel therapeutic approach for managing PKD, which currently has limited treatment options. The study aims to ensure that the gene therapy is safe for patients, which is a critical step before further efficacy studies can be conducted.
Participants
The clinical trial involves a total of **five participants** diagnosed with **Pyruvate Kinase Deficiency**, an inherited metabolic disorder affecting red blood cell survival. The study population includes both male and female subjects, with an age range divided into two cohorts: adults aged 18 to 50 years and pediatric participants aged 8 to less than 18 years. Participants were selected based on a confirmed PKLR mutation and a history of severe and/or transfusion-dependent anemia. The trial includes individuals with adequate cardiac, pulmonary, renal, and hepatic function, as well as those with detailed medical records of transfusion requirements over the past two years. Both vulnerable populations and those capable of providing informed consent or assent are included. Lifestyle factors such as diet and physical activity are not specified in the available data.
Plans and Procedures
The clinical trial is designed to evaluate the safety and toxicity of **autologous CD34+ hematopoietic stem cells** transduced with a lentiviral vector encoding the codon-optimized version of the PKLR gene in subjects with **Pyruvate Kinase Deficiency**. This is a Phase I trial, characterized by a randomized, double-blind, controlled design, ensuring that neither the participants nor the researchers know who is receiving the investigational product or the control. The trial is expected to last until October 2025, with recruitment having started in January 2020.
Participants will undergo a series of study visits, beginning with an inclusion (screening) visit to confirm eligibility based on criteria such as a confirmed PKLR mutation and a history of severe anemia. The trial includes both adult and pediatric cohorts, with specific age and health criteria for each group. Following the screening, participants will receive the investigational product through **intravenous infusion**. Subsequent follow-up visits will monitor safety, toxicity, and the achievement of stem cell engraftment, defined by specific neutrophil and platelet counts. The end-of-study visit will assess the primary and secondary endpoints, including insertional mutagenesis and immunogenicity.
Participant involvement is expected to last for the duration of the trial, with the possibility of early termination if adverse events occur or if the participant withdraws consent. Conditions such as inadequate cardiac, pulmonary, renal, or hepatic function may also lead to early termination. The trial aims to provide comprehensive data on the safety profile of the investigational product, contributing to the understanding of its potential therapeutic benefits for individuals with Pyruvate Kinase Deficiency.
Treatment
The clinical trial involves the administration of several treatments, including **Busulfan Fresenius Kabi 6 mg/ml concentrate for solution for infusion**. This experimental medication is a **concentrate for solution for infusion** and is administered via **infusion**. The active substance in this product is **busulfan**, a chemical compound. The dosage is calculated based on body weight, with a maximum daily dose of 4 mg/kg and a total maximum dose of 16 mg/kg over a treatment period of up to 4 days. This medication is used as a conditioning treatment prior to hematopoietic stem cell transplantation (HSCT).
Another treatment used in the trial is **Neupogen 30 MU (0.6 mg/ml) solución inyectable en jeringa precargada filgrastim**. This product is a **solution for injection/infusion** and is administered via **subcutaneous injection**. The active substance is **filgrastim**, a protein derived from genetic recombination. The dosage is also weight-based, with a maximum daily dose of 10 µg/kg and a total maximum dose of 70 µg/kg over a treatment period of up to 7 days. This medication is used to mobilize hematopoietic stem cells to peripheral blood for collection.
**Mozobil 20 mg/ml solution for injection** is another treatment included in the study. It is a **solution for injection** administered via **subcutaneous injection**. The active substance is **plerixafor**, a chemical compound. The dosage is weight-based, with a maximum daily dose of 240 µg/kg and a total maximum dose of 720 µg/kg over a treatment period of up to 3 days. This medication is used to enhance the mobilization of hematopoietic stem cells to peripheral blood for collection.
The primary investigational product in this trial is **Merilen**, which is an **infusion** administered via **intravenous infusion**. The active substance consists of **autologous CD34+ hematopoietic stem cells transduced ex vivo with a lentiviral vector encoding the codon-optimized version of the PKLR gene**. This gene therapy product is administered as a single dose with a maximum total dose of 4,000,000 cells. The treatment is designed to address pyruvate kinase deficiency (PKD) by introducing a functional version of the PKLR gene into the patient's hematopoietic stem cells.
Efficacy
Efficacy in this clinical trial will be assessed through both primary and secondary endpoints. The primary endpoints include the evaluation of **insertional mutagenesis**, which involves assessing the gene-modified clonal repertoire and lentiviral insertion site analysis (ISA) in blood and, if feasible, bone marrow cells via modified gene sequencing (MGS)-PCR. Additionally, the presence of replication-competent lentivirus (RCL) will be evaluated in blood, particularly in cases where there is clinical suspicion of unexplained viral illness. Immunogenicity will also be assessed by detecting antibodies against the codon-optimized red cell pyruvate kinase (coRPK) or other lentiviral vector components in blood serum.
Secondary endpoints focus on the achievement and timing of stem cell engraftment, defined as neutrophil engraftment occurring within 42 days post-infusion. Neutrophil engraftment is characterized by an absolute neutrophil count of ≥500/μL for three consecutive days. Platelet engraftment is defined as a platelet count of ≥20,000/μL for three consecutive days, independent of thrombopoietin mimetics and platelet transfusions for at least seven days. The incidence of respiratory complications will also be monitored as part of the secondary efficacy assessments.
Inclusion and Exclusion Criteria
Inclusion Criteria
- PKD diagnosis with a confirmed PKLR mutation. 2. Patient age at enrollment as follows: a. Adult Cohort (n=2): Age ≥18 years and ≤50 years. b. Pediatric Cohort (n=2-3): Age ≥8 to <18 years 3. History of severe and/or transfusion-dependent anemia, defined as: a. At least 6 RBC transfusion episodes over a prior 12-month period and Hb levels <9.5 g/dL in the previous 1 months despite prior splenectomy OR b. At least 3 RBC transfusion episodes per year over 2 prior years and Hb levels <9.5 g/dL in the previous 12 months despite prior splenectomy OR c. Hb levels <8.0 g/dL despite prior splenectomy in the absence of transfusions (documented during 2 or more assessments during the prior 1–-2 years) regardless of transfusion requirements. 4. Adequate cardiac, pulmonary, renal and hepatic function, as detailed in relevant exclusion criteria. 5. Availability of detailed medical records, including transfusion requirements, for at least the prior 2 years. 6. Willing and able to read and correctly understand the patient information sheet and provide consent (or informed assent for minors) regarding study participation. 7. Negative serum pregnancy test for female patients of childbearing potential.
Exclusion Criteria
- Presence of other known causes of hemolysis (in addition to PKD). Patients with concurrent G6PD deficiency diagnosed during pre-study evaluation may be considered for eligibility if in the opinion of the Investigator, the hemolytic anemia is the result of PKD and the G6PD deficiency is considered an incidental finding. 2. A venous thromboembolic event (VTE; i.e., pulmonary embolism or deep vein thrombosis) or arteriothromboembolic event (ATE; including unstable angina, myocardial infarction, stroke or transient ischemic attack) during the prior 12 months. 3. Any evidence of severe iron overload that, per Investigator discretion, warrants exclusion. 4. Evidence of bridging fibrosis, cirrhosis or active hepatitis on liver biopsy. Liver biopsy is required when liver iron concentration (LIC) is ≥ 15 mg/g on T2* magnetic resonance imaging (MRI) of liver. If a liver biopsy has been performed less than 6 months prior to enrollment, it does not need to be repeated. 5. Significant medical conditions including documented HIV infection, active viral hepatitis, poorly-controlled hypertension, pulmonary hypertension, cardiac arrhythmia or congestive heart failure; or ATEs (including stroke or myocardial infarction) within the 6 prior months. 6. Active hematologic or solid organ malignancy, not including nonmelanoma skin cancer or another carcinoma in situ. Patients with previously resected solid organ malignancies or definitively treated hematologic malignancies may be eligible if there has been no evidence of active malignancy during the prior 3 years. 7. Uncontrolled seizure disorder. 8. Cardiac T2* <10 ms by magnetic resonance imaging (MRI) or left ventricular ejection fraction (LVEF) <45% by echocardiogram or multiple gated acquisition scan (MUGA). 9. Hepatic dysfunction as defined by: • Alanine aminotransferase (ALT) or aspartate aminotransferase (AST) >2.5 × the upper limit of normal (ULN). 10. Renal dysfunction defined as serum creatinine >ULN. Patients with creatinine above ULN may be eligible pending documentation of a glomerular filtration rate ≥ 60 mL/min/1.73m2 as calculated by Modification of Diet in Renal Disease equation, the revised Schwartz formula (for patients under 18 years old), or 24-hour urine collection. 11. Pulmonary dysfunction as defined by either: • Need for supplemental oxygen during the prior 2 weeks (in absence of acute infection) or • Oxygen saturation (by pulse oximetry) <90%. 12. Any medical or other contraindication for leukapheresis as determined by the treating Investigator. 13. Any medical or psychiatric condition that in the opinion of the Investigator renders the patient unfit for trial participation or at higher than acceptable risk for participation. 14. Poor functional status, evidenced by Karnofsky Index <70 in adults or Lansky <70 in children. 15. Participation in another clinical trial with an investigational drug within 14 days before the informed consent signature. Participation in observational studies is allowed. 16. Pregnant women or women with a positive serum pregnancy test at screening or breast feeding or planning to become pregnant within the next 24 months. Women not willing to use highly effective contraceptive methods during the complete study period.* 17. Previous allogeneic or other hematopoietic stem cell transplant * Females of childbearing age potential and male patients with partners of childbearing potential must use highly effective contraceptive measures (according to Clinical Trials Facilitation Group recommendations).
Trial Status by Country
| Country | Status | Start of Recruitment | Planned Patients |
|---|---|---|---|
 Spain | Not Recruiting | 13 Jan 2020 | 3 |
Sites & Investigators
Research sites
Investigators
Investigational Products
Details about the medicinal products being studied in this clinical trial.
| Product Name | Role in Trial | Formulation | Administration | Max Daily Dose | Treatment Duration | EU MP Number |
|---|---|---|---|---|---|---|
Busulfan Fresenius Kabi 6 mg/ml concentrate for solution for infusion | Other | CONCENTRATE FOR SOLUTION FOR INFUSION | INFUSION | 4 | 4 | PRD1938253 |
Neupogen 30 MU (0,6 mg/ml) solución inyectable en jeringa precargada filgrastim | Other | SOLUCIÓN INYECTABLE EN JERINGA PRECARGADA | SUBCUTANEOUS INJECTION | 10 | 7 | PRD729930 |
Mozobil 20 mg/ml solution for injection | Other | SOLUTION FOR INJECTION | SUBCUTANEOUS INJECTION | 240 | 3 | PRD382671 |
Merilen | Test | INFUSION | INTRAVENOUS INFUSION | 4000000 | 1 | PRD7873153 |