Gene Therapy for Adenosine Deaminase Severe Combined Immune Deficiency Using Peripheral Blood and EFS ADA Vector

Inclusion Criteria:

All subjects must fulfill the following criteria to be included in the study:

1. Provision of written informed consent prior to any study related procedures. In thisstudy consent must be provided by the parents/legal guardians and, where applicableaccording to local laws, a signed assent from the child,

2. Subjects ≥30 days of age,

3. With a diagnosis of ADA-SCID based on: Evidence of ADA deficiency, defined as: i. Decreased ADA enzymatic activity in erythrocytes, leukocytes, skin fibroblasts,or in cultured fetal cells to levels consistent with ADA-SCID as determined by thereference laboratory, or ii. Identified mutations in ADA alleles consistent with asevere reduction in ADA activity, Evidence of ADA-SCID based on either: i. Family history of a first order relative with ADA deficiency and clinical andlaboratory evidence of severe immunologic deficiency, or ii. Evidence of severeimmunologic deficiency in subjects prior to the institution of immune restorativetherapy, based on

4. Lymphopenia (absolute lymphocyte count (ALC) <400 cells/mL) OR absence or lownumber of T cells (absolute CD3+ count < 300 cells/mL), or

5. Severely decreased T lymphocyte blastogenic responses to phytohemagglutinin (either <10% of lower limit of normal controls for the diagnostic laboratory,or <10% of the response of the normal control of the day, or stimulation index <10), or

6. Identification of SCID by neonatal screening revealing low T Cell ReceptorExcision Circles (TREC) levels.

7. Ineligible for matched family allogeneic bone marrow (BM) transplantation, definedas the absence of a medically eligible HLA-identical sibling or family donor, withnormal immune function, who could serve as an allogeneic bone marrow donor.

8. Females of child-bearing age will be required to provide a negative pregnancy test 30 days prior to Visit 2.

9. Subjects and their parents/legal guardians must be willing and able to comply withstudy restrictions and to remain at the clinic for the required duration during thestudy period and willing to return to the clinic for the follow up evaluation asspecified in the protocol.

Exclusion Criteria:

Subjects will not be eligible for the study if any of the following criteria is fulfilled:

1. Ineligible for autologous HSCT as per clinical site criteria

2. Other conditions which in the opinion of the Principal Investigator and/or CoInvestigators, contraindicate the mobilization of peripheral blood or theleukapheresis process, the administration of busulfan and the infusion of transducedcells, or which indicate an inability of the subject or subject's parent/legalguardian to comply with the protocol

3. Hematologic abnormality, defined as:

• Anemia (Hb <8.0 g/dl).

• Neutropenia (ANC <500/mm3). Note: ANC <500 with absence of myelodysplasticsyndrome on bone marrow aspirate and biopsy and normal marrow cytogenetics areacceptable for eligibility.

• Thrombocytopenia (platelet count <50,000/mm3, at any age).

• Prothrombin time or international normalized ratio (INR) and partialthromboplastin time (PTT) >2 x upper limit of normal (ULN) (subjects with acorrectable deficiency controlled on medication will not be excluded).

• Cytogenetic abnormalities on peripheral blood or bone marrow or amniotic fluid (if available).

• Prior allogeneic HSCT with cytoreductive conditioning.

4. Pulmonary abnormality, defined as:

• Resting O2 saturation by pulse oximetry <90% on room air.

• Chest X-ray indicating active or progressive pulmonary disease. Note: Chest Xray indicating residual signs of treated pneumonitis is acceptable foreligibility.

5. Cardiac abnormality, defined as:

• Abnormal ECG indicating cardiac pathology.

• Uncorrected congenital cardiac malformation with clinical symptoms.

• Active cardiac disease, including clinical evidence of congestive heartfailure, cyanosis, hypotension.

• Poor cardiac function as evidenced by left ventricular ejection fraction <40%on echocardiogram.

6. Neurologic abnormality, defined as:

• Significant neurologic abnormality revealed by examination.

• Uncontrolled seizure disorder.

7. Renal abnormality, defined as:

• Renal insufficiency: serum creatinine ≥1.2 mg/dl (106 µmol/L), or ≥3+proteinuria.

• Abnormal serum sodium, potassium, calcium, magnesium or phosphate levels at >2x ULN.

8. Hepatic/gastrointestinal abnormality, defined as:

• Serum transaminases >5 x ULN.

• Serum bilirubin >2 x ULN.

• Serum glucose >1.5 x ULN.

9. Oncologic disease, defined as:

• Evidence of active malignant disease other than dermatofibrosarcoma protuberans (DFSP).

• Evidence of DFSP expected to require anti-neoplastic therapy within the 5 yearsfollowing the infusion of genetically corrected cells (if anti-neoplastictherapy has been completed, a subject with a history of DFSP can be included).

• Evidence of DFSP expected to be life limiting within the 5 years following theinfusion of genetically corrected cells.

10. Known sensitivity to Busulfan.

11. Confirmation of an infectious disease by deoxyribonucleic acid (DNA) polymerasechain reaction (PCR) positive at time of assessment for the following:

• HIV-1,

• Hepatitis B,

• Parvovirus B19.

12. The subject is pregnant or has a major congenital anomaly.

13. Is likely to require treatment during the study with drugs that are not permitted bythe study protocol.

14. The subject has previously received another form of gene therapy.