Fludarabine-based Conditioning for Severe Aplastic Anemia (BMT CTN 0301)

• Graft Failure - defined by lack of neutrophil engraftment (ANC less than 0.5 x 10^9/L for 3 consecutive days on different days) [ Time Frame: Day 100 ] [ Designated as safety issue: Yes ]
• Regimen-related Toxicity (RRT) - scored according to the Bearman scale [ Time Frame: Day 100 ] [ Designated as safety issue: Yes ]
• Early Death [ Time Frame: Day 100 ] [ Designated as safety issue: Yes ]

• Primary Graft Failure - defined by lack of neutrophil engraftment (ANC < 0.5 x 109/L for 3 consecutive days on different days) by 42 days post-transplant
• Regimen-related Toxicity (RRT) - scored according to the Bearman scale
• Early Death - defined as death prior to Day 42 post-transplant

• Secondary Graft Failure - defined by initial neutrophil engraftment followed by subsequent decline in the ANC to less than 0.5 x 10^9/L for three consecutive measurements on different days, unresponsive to growth factor [ Time Frame: Day 100 ] [ Designated as safety issue: No ]
• Acute GVHD of Grades 2-4 and 3-4 - graded according to the BMT CTN Manual of Procedures [ Time Frame: Day 100 ] [ Designated as safety issue: No ]
• Chronic GVHD - Chronic GVHD is scored according to the BMT CTN MOP. The first day of chronic GVHD onset will be used to calculate cumulative incidence curves. [ Time Frame: Day 730 ] [ Designated as safety issue: No ]
• Post-transplant Survival [ Time Frame: Day 100 ] [ Designated as safety issue: Yes ]

• Secondary Graft Failure - defined (in patients surviving at least 42 days) by initial neutrophil engraftment followed by subsequent decline in the ANC to < 0.5 x 109/L for three consecutive measurements on different days, unresponsive to growth factor
• Acute GVHD of Grades 2-4 and 3-4 - graded according to the BMT CTN Manual of Procedures (measured through 100 days)
• Chronic GVHD - Chronic GVHD is scored according to the BMT CTN MOP. The first day of chronic GVHD onset will be used to calculate cumulative incidence curves.

The purpose of the current study is to continue to optimize conditioning regimens in high-risk patients with severe aplastic anemia transplanted with marrow from HLA-compatible unrelated donors. Specifically, the study will determine whether the addition of fludarabine to the conditioning regimen previously described by Deeg et al. will permit a reduction in the CY dose, to a point where sustained hematopoietic engraftment and survival are maintained (or improved), while the frequency of major regimen-related toxicity (RRT) and early deaths is reduced.

BACKGROUND:

Aplastic anemia (AA) remains a life-threatening illness. Treatment options include supportive care (transfusions, growth factors, etc.), immunosuppression therapy and stem cell transplantation. Only the latter two have favorably impacted the natural history of the disease. The prognosis of AA patients, particularly SAA, as defined by Camitta et al., who fail to respond to immunosuppressive therapy (IS) or who relapse after an initial response to IS is poor. Although many of these patients can be supported in the short term with growth factors, transfusions and possibly rechallenged successfully with IS, the cumulative morbidity and mortality from infection, hemorrhage or transfusion-related complications is substantial.

While allogeneic bone marrow transplantation is potentially curative in AA, no more than 25% of patients have an HLA-identical sibling donor. Cyclophosphamide (CY)-ATG has been recommended as the preparative regimen of choice in sibling donor transplants. Results of bone marrow transplantation from alternative donors, such as matched unrelated donors and mismatched related donors in AA patients who have failed IS, have largely been unsatisfactory. The cyclophosphamide-ATG conditioning regimen has proved inadequate in ensuring engraftment in allogeneic transplants from matched, unrelated donors for AA. This was the major reason why total body radiation (TBI) has been added to the conditioning regimen.

Graft failure is a very serious and frequently life-threatening or fatal event following MUD allografts in aplastic anemia. It is an immunologically mediated event. Risk factors for graft failure include the use of HLA nonidentical or unrelated donors, a poor marrow nucleated cell dose as well as prolonged transfusional support prior to BMT (which increases the probability of patient sensitization to multiple antigens). While some patients may achieve autologous hematopoietic recovery, prolonged pancytopenia is common and infection-related morbidity and mortality are very substantial. Reconditioning for a second allograft from the same or a different donor is frequently not successful. While the addition of TBI and intensive pre-transplant conditioning has led to a sizable improvement in engraftment rates, this has come with a price, particularly in adult patients. Transplant-related toxicity has been a major and frequent problem. Radiation-induced pulmonary toxicity in particular has been common, usually in the form of diffuse alveolar damage or diffuse interstitial pneumonitis. In addition, GVHD-related morbidity and mortality in these patients have also been substantial.

DESIGN NARRATIVE:

The study is a prospective Phase I/II dose optimization study. All patients are given a fixed dose of ATG (either thymoglobulin: 3 mg/kg IV daily x 3 or ATGAM 30 mg/kg IV daily x 3, on Days -4 to -2), Fludarabine (30 mg/m^2 IV daily x 4, on Days - 5 to -2), and TBI (200 cGy from a linear accelerator at less than 20 cGy/min on Day -1). The starting CY dose will be 150 mg/kg (50 mg/kg intravenously daily, Days -4 to -2), and will be de-escalated depending on engraftment and toxicity. The Phase I portion of the trial (maximum of 24-27 patients) tests each of four dose levels of CY for adequate safety and graft retention. The Phase II portion of the trial refines the dose selection and allocates an additional 54 patients to the optimal dose, at which two-year post-transplant survival will be assessed. The combined enrollment in Phase I and II will total 78-81 patients.

The study is a prospective single-arm Phase I/II dose-selection and evaluation study. The study will seek the optimal dose level of CY based on assessments of graft failure, toxicity and early death during 100 days of follow-up post-transplant. A brief synopsis is given below.

Phase I - Test Each Dose for Adequate Safety and Graft Retention

1. Proceed from the highest dose (150 mg/kg CY) to the lowest dose (0 mg/kg CY), treating a minimum of six patients at each dose.
2. Evaluate the 100-Day outcomes for toxicity, death and graft failure on each patient enrolled at the current dose, or until stopping criteria are met.
3. If there are three or more graft failures at the current dose, the current dose and all lower doses are closed to further enrollment.
4. If there are five or more severe regimen-related toxicities and/or early deaths at the current dose, the current dose is closed to further enrollment, and the next lower dose is tested.
5. Dose de-escalation ceases once all four doses are tested or closed to further enrollment.

Phase II - Refine Dose Selection and Allocate Patients to the Optimal Dose

1. Treat each newly enrolled patient at the most desirable of the dose levels remaining open to enrollment. This can involve de-escalation, escalation, or no change in dose.
2. As each patient completes the observation period, evaluate the 100-Day outcomes for graft failure, toxicity and/or early death for this patient, or until stopping criteria are met.
3. If there are excess (according to the criteria in Table 5.8) graft failures, that patient's dose and all lower doses are closed to further enrollment.
4. If there are excess (according to the criteria in Table 5.8) toxicities and/or early deaths, that patient's dose is closed to further enrollment.
5. Re-evaluate the desirability of the current dose level based on the 100-Day outcomes for toxicity and/or early death and graft failure.
6. Repeat steps 1-5 until 54 patients are enrolled in Phase II, or all dose levels are closed to further enrollment.

Dosage Levels for CY:

3 Days (Day -4, -3, -2): Dose of 50 mg/kg/day; total dose of 150 mg/kg; dose level 3

2 Days (Day -3, -2): Dose of 50 mg/kg/day; total dose of 100 mg/kg; dose level 2

1 Day (Day -2): Dose of 50 mg/kg/day; total dose of 50 mg/kg; dose level 1

0 Days (None): No dose; no total dose; dose level 0

There may be wait periods between enrollment of successive patients and/or cohorts for endpoint assessment. Under these circumstances, the final decision about waiting versus treating the patient off study will be made at the local transplant center.

Primary Outcomes:

1. Graft Failure: Neutrophil engraftment is defined as the achievement of an ANC ≥ 0.5 x 10^9/L for three consecutive measurements on different days. Primary graft failure is defined by the lack of neutrophil engraftment; i.e., ANC < 0.5 x 10^9/L measured for three consecutive measurements on different days by 100 days post-transplant. Secondary graft failure prior to Day 100 post-transplant will count towards the graft failure endpoint.
2. Regimen-related Toxicity (RRT): RRT will be scored according to the Bearman scale. Major RRT is defined as severity of grade 4 in any organ system or grade 3 for pulmonary, cardiac, renal, oral mucosal or hepatic, in keeping with the approach adopted in FHCRC Protocol #800. The assessment for RRT will be carried out weekly until Day 100 post-transplant. The NCI's CTCAE version 3.0 will be used to supplement the Bearman toxicity criteria.
3. Early Death:This endpoint is defined as death prior to Day 100 post-transplant.

Secondary Outcomes:

1. Post-transplant survival- as defined as time from transplant to death from any cause.
2. Secondary Graft Failure - This endpoint is defined (in patients surviving at least 100 days) by initial neutrophil engraftment followed by subsequent decline in the ANC to < 0.5 x 10^9/L for 3 consecutive measurements on different days, unresponsive to growth factor therapy.
3. Acute GVHD of Grades 2-4 and 3-4 - Acute GVHD is graded according to the Blood and Marrow Transplant Clinical Trials Network (BMT CTN) Manual of Procedures (MOP). The first day of acute GVHD onset at a certain grade will be used to calculate cumulative incidence curves for that GVHD grade (e.g., if the onset of grade 1 acute GVHD is on Day 19 post-transplant and onset of grade 3 is on Day 70 post-transplant, time to grade 3 is Day 70). This endpoint will be evaluated through 100 days.
4. Chronic GVHD - Chronic GVHD is scored according to the BMt CTN MOP. The first day of chronic GVHD onset will be used to calculate cumulative incidence curves.

• Drug: Cyclophosphamide
  total dose 150 mg/kg given as 50 mg/kg per day on Days -4, -3, -2
• Drug: Cyclophosphamide
  total dose 100 mg/kg given as 50 mg/kg per day on Days -3, -2
• Drug: Cyclophosphamide
  total dose 50 mg/kg given as 50 mg/kg per day on Day -2
• Drug: Cyclophosphamide
  total dose 0 mg/kg

• Experimental: 1 Cyclophosphamide
  150 mg/kg Cyclophosphamide (total dose)
  Intervention: Drug: Cyclophosphamide
• Experimental: 2 Cyclophosphamide
  100 mg/kg Cyclophosphamide (total dose)
  Intervention: Drug: Cyclophosphamide
• Experimental: 3 Cyclophosphamide
  50 mg/kg Cyclophosphamide (total dose)
  Intervention: Drug: Cyclophosphamide
• Experimental: 4 Cyclophosphamide
  0 mg/kg Cyclophosphamide (total dose)
  Intervention: Drug: Cyclophosphamide

Inclusion Criteria:

• Patients up to 65 years of age at time of registration with a diagnosis of SAA; SAA is defined as follows:

  1. Bone marrow cellularity less than 25% or marrow cellularity less than 50% but with less than 30% residual hematopoietic cells
  2. Two out of three of the following (in peripheral blood): neutrophils less than 0.5 x 10^9/L; platelets less than 20 x 10^9/L; reticulocytes less than 20 x 10^9/L
• Patient must have an available unrelated donor with a 7/8 or 8/8 match for HLA-A, B, C, and DRB1 antigen; typing is by DNA techniques: intermediate resolution for A, B, and C, and high resolution for DRB1; HLA-DQ typing is recommended but will not count in the match
• Patient and/or legal guardian able to provide signed informed consent
• Matched unrelated donor must consent to provide a marrow allograft

• Patients with adequate organ function as measured by:

  1. Cardiac: left ventricular ejection fraction at rest must be greater than 40% or shortening fraction greater than 20%
  2. Hepatic: serum bilirubin less than 2x upper limit of normal for age as per local laboratory) (with the exception of isolated hyperbilirubinemia due to Gilbert's syndrome), ALT and AST less than 4x upper limit of normal for age (as per local laboratory)
  3. Renal: serum creatinine less than 2x upper limit of normal for age (as per local laboratory)
  4. Pulmonary: FEVl, FVC, and DLCO (corrected for Hb) greater than 50% predicted; for patients in which pulse oxymetry is performed, O2 saturation greater than 92%
• Diagnosis of Fanconi anemia must be excluded in patients younger than 18 years of age by diepoxybutane testing on peripheral blood or comparable testing on marrow.

Exclusion Criteria:

• Clonal cytogenetic abnormalities associated with MDS or AML on marrow examination
• Diagnosis of other "congenital" aplastic anemias such as: Diamond-Blackfan; Shwachmann-Diamond; congenital amegakaryocytosis
• Symptomatic or uncontrolled cardiac failure or coronary artery disease
• Karnofsky performance status less than 60% or Lansky less than 40% for patients younger than 16 years old
• Uncontrolled bacterial, viral or fungal infections (currently taking medication and progression of clinical symptoms)
• Seropositive for the human immunodeficiency virus (HIV)
• Pregnant (positive total HCG) or breastfeeding
• Presence of large accumulation of ascites or pleural effusions, which would be a contraindication to the administration of methotrexate for GVHD prophylaxis
• Known severe or life-threatening allergy or intolerance to ATG or cyclosporine/ tacrolimus
• Planned administration of alemtuzumab (Campath-1H) or other investigational agents as alternative agent for GVHD prophylaxis
• Concomitant enrollment in a Phase I study
• Positive patient anti-donor lymphocyte crossmatch in HLA-A or B mismatched transplants; the definition of match is in Section 2.2.1; the crossmatch would only apply to mismatches at HLA-A or B, not DRB1 or HLA-C
• Prior allogeneic marrow or stem cell transplantation
• Patients with prior malignancies except resected basal cell carcinoma or treated carcinoma in-situ; cancer treated with curative intent less than 5 years previously will not be allowed unless approved by the Medical Monitor or Protocol Chair; cancer treated with curative intent more than 5 years previously will be allowed

• National Heart, Lung, and Blood Institute (NHLBI)
• Blood and Marrow Transplant Clinical Trials Network
• National Cancer Institute (NCI)