Gene Therapy for Pyruvate Kinase Deficiency (PKD)

Inclusion Criteria:

   - PKD diagnosis with a confirmed PKLR mutation.

   - Adult Cohort ≥18 years old and <50 years for the initial 2 patients enrolled;
   Pediatric Cohort ≥8-17 years for the next 2-3 patients.

   - History of severe, transfusion-dependent anemia, defined as:

      1. At least 6 red blood cell transfusion episodes over a prior 12-month period and
      Hb levels <9.5 g/dL in the previous 12 months despite splenectomy OR

      2. At least 3 red blood cell transfusion episodes per year over 2 prior years (in
      the absence of precipitating events such as infection or surgery) and Hb levels
      <9.5 g/dL in the previous 12 months despite prior splenectomy.

      OR

      3. 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.

   - Adequate cardiac, pulmonary, renal and hepatic function, as detailed in relevant
   exclusion criteria.

   - Availability of detailed medical records, including transfusion requirements, for at
   least the prior 2 years.

   - Willing and able to read and correctly understand the patient information sheet and
   provide consent (or informed assent for minors) regarding study participation.

   - 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.

   - 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.

   - Any evidence of severe iron overload that, per Investigator discretion, warrants
   exclusion.

   - 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.

   - Significant medical conditions including documented HIV infection, active viral
   hepatitis, poorly-controlled hypertension, pulmonary hypertension cardiac arrhythmia
   or congestive heart failure; pulmonary hypertension or ATEs (including stroke or
   myocardial infarction) within the 6 prior months.

   - Active hematologic or solid organ malignancy, not including non-melanoma 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.

   - Uncontrolled seizure disorder.

   - Cardiac T2*<10 ms by magnetic resonance imaging (MRI) or left ventricular ejection
   fraction (LVEF) <45% by echocardiogram or multiple gated acquisition (MUGA) scanning.

   - Hepatic dysfunction as defined by: alanine aminotransferase (ALT) or aspartate
   aminotransferase (AST) >2.5× the upper limit of normal.(ULN).

   - Renal dysfunction as defined as serum creatinine > ULN. Patients with creatinine above
   ULN may be eligible pending documentation of a GFR ≥60 mL/min/1.73m2 as calculated by
   the Modification of Diet in Renal Disease equation (Stevens 2006), the revised
   Schwartz formula (for patients under 18 years old) (Schwartz 2009), or 24-hour urine
   collection.

   - 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%.

   - Any medical or other contraindication for both leukapheresis and BM harvest procedure
   as determined by the treating Investigator.

   - Any medical or psychiatric condition that in the opinion of the Investigator renders
   the subject unfit for trial participation or at higher than acceptable risk for
   participation.

   - Poor functional status, evidenced by Karnofsky Index <70 in adults or Lansky <70 in
   children.

   - Participation in another clinical trial with an investigational drug within 14 days
   before the informed consent signature. Participation in observational studies is
   allowed.

   - 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.