Methods to Enhance the Safety and Effectiveness of Stem Cell Transplants
Methods to Enhance the Safety and Effectiveness of Stem Cell Transplants
Bone marrow stem cell transplants (otherwise called bone marrow transplants) from healthy
 donors are sometimes the only means of curing hematological malignant diseases such as acute
 and chronic leukemias, myelodysplastic syndrome, myeloproliferative diseases and lymphomas.
 Before transplant the patient receives chemotherapy and radiation treatment to reduce the
 malignancy to low levels and to prevent rejection of the transplant. The transplant restores
 the blood counts to normal and replaces the patients immunity with that of the donor. The
 donor's immune cells increase the effect of the transplant by attacking remaining malignant
 cells. Donor immune cells (especially those called T lymphocytes) also attack healthy
 non-cancerous cells and tissues of the recipient causing "graft-versus-host-disease" (GVHD).
 Strong GVHD reactions occurring within weeks after the transplant can be life-threatening .
 In this study we remove most of the T lymphocytes from the transplant to minimize the risk of
 GVHD. However to improve immunity against residual malignant cells and boost immunity to
 infections, donor T cells (stored frozen at time of transplant) are given back around 90 days
 after the transplant when they have a reduced risk of causing serious GVHD.

 Any patient between 10 and 75 years of age with acute or chronic leukemia, myelodysplastic
 syndrome, myeloproliferative syndromes or lymphoma, who have a family member who is a
 suitable stem cell donor may be eligible for this study. Candidates are screened with a
 medical history and various tests and examinations.
Peripheral blood stem cell transplant research carried out by the National Heart Lung and
 Blood Institute (NHLBI) Bonne Marrow Transplantation (BMT) Unit focus on transplant
 techniques designed to decrease graft versus host disease (GVHD), increase the
 graft-versus-leukemia (GVL) effect and reduce the risk of post-transplant graft rejection.

 We have found that by controlling the stem cell (CD34+ cell) and T lymphocyte (CD3+ cell)
 dose severe GVHD can be reduced whilst beneficial GVL effects can be preserved by postponed
 donor lymphocyte infusion (DLI). We found that T cell depleted transplants using the
 Nexell/Baxter Isolex 300i system to obtain high CD34+ doses depleted of lymphocytes were safe
 to administer and associated with less severe acute GVHD and promising response rates and
 overall survival, when combined with a delayed T cell add-back (DLI). This protocol is
 designed to evaluate safety and efficacy of an improved T cell depletion procedure using the
 Miltenyi CliniMACs system (CD34 reagent system) with a delayed T cell add back in the
 HLA-matched peripheral blood stem cell transplant setting.

 The protocol will accrue up to 68 subjects ages 10-75 with a hematological malignancy, in
 whom allogeneic stem cell transplantation from an HLA-matched sibling would be routinely
 indicated. 68 sibling donors will also be recruited. Diagnostic categories will include acute
 and chronic leukemia, myelodysplastic syndrome, lymphoma, multiple myeloma and
 myeloproliferative syndromes.

 Subjects will receive a myeloablative conditioning regimen of cyclophosphamide, fludarabine
 and total body irradiation, followed by an infusion of a stem cell product prepared using the
 Miltenyi CliniMacs system for CD34 selection, and a delayed T-cell add back as donor
 lymphocyte infusion (DLI) at day 90. Older subjects will receive a lower dose of irradiation
 to reduce the regimen intensity.

 Participants undergo apheresis to collect lymphocytes for research studies. This procedure
 involves collecting blood through a needle in the arm, extracting the lymphocytes through a
 cell separator machine and returning the rest of the blood through a needle in the other arm.
 Before treatment begins, patients have a central intravenous line (plastic tube) placed in a
 vein in the neck. This line remains in place during the transplant and recovery period for
 drawing and transfusing blood, giving medications, and infusing the donated cells.

 To prepare patients for transplantation their immune system is suppressed by a combination of
 chemotherapy and radiation which will also help to fight remaining malignant cells. The
 chemotherapy consists of two anti-cancer drugs (fludarabin and cyclophosphamide) and will be
 infused over the central line starting eight days before the transplant. Patients will
 receive eight doses of total body irradiation, administered in two 30-minute sessions per day
 for 4 days. Patients above 55 years of age who tolerate transplants less well than younger
 individuals will receive a lower dose of radiation. One day after the preparative treatment
 is finished, patients receive the transplant of donors' stem cells as an infusion through the
 central line.

 Patients receive cyclosporine, an immune-suppressing drug starting 6 days before the
 transplant until 21 days post-transplant. This helps prevent both transplant rejection and
 GVHD. The drug is stopped to allow the donor immunity to function and is restarted on day 89
 to prevent GVHD from the infusion of T lymphocytes on day 90.

 Patients are followed with various tests, treatments and examinations periodically for the
 first 3 years and then yearly thereafter.

 Primary endpoint will be overall survival at day +200. Non-relapse mortality at day +200 will
 be monitored for safety. Secondary endpoints will be standard transplant outcome variables
 such as non-hematologic toxicity, incidence and severity of acute and chronic GVHD and
 relapse of disease.
INCLUSION CRITERIA - RECIPIENT:

 -  Ages 10-75 years inclusive

 -  Chronic myelogenous leukemia (CML):

 -  Subjects under the age of 21 in chronic phase

 -  Subjects ages 10-75 in chronic phase who have failed treatment with imatinib, have
 intolerance to imatinib, or who did not receive imatinib at therapeutic doses within
 the first 12 months from diagnosis.

 -  Subjects ages 10-75 in accelerated phase or blast transformation.

 -  Acute lymphoblastic leukemia (ALL): any of these categories: ALL in first remission
 with high-risk features (presenting leukocyte count greater than 100,000/cu mm,
 Karyotypes t9; 22, t4, t19, t11, biphenotypic leukemia) All second or subsequent
 remissions, primary induction failure, partially responding or untreated relapse.

 -  Acute myelogenous leukemia (AML): AML in first remission - except AML with good risk
 karyotypes: AML M3 (t15; 17), AML M4Eo (inv 16), AML t (8; 21). All AML in second or
 subsequent remission, primary induction failure and resistant relapse

 -  Myelodysplastic syndromes (MDS): any of these categories - refractory anemia with
 transfusion dependence, refractory anemia with excess of blasts, transformation to
 acute leukemia, chronic myelomonocytic leukemia, atypical MDS/myeloproliferative
 syndromes

 -  Myeloproliferative disorders including atypical (Ph negative) chronic myeloid and
 neutrophilic leukemias, progressing myelofibrosis, and polycythemia vera, essential
 thrombocythemia in transformation to acute leukemia or with progressive transfusion
 requirements or pancytopenia.

 -  Chronic lymphocytic leukemia refractory to fludarabine treatment and with bulky
 progressive disease or with thrombocytopenia (less than or equal to 100,000 /microl)
 or anemia (less than or equal to 10g/dl) not due to recent chemotherapy.

 -  Non-Hodgkins lymphoma including Mantle cell lymphoma relapsing or refractory to
 standard of care treatments

 -  Multiple myeloma, Waldenstroms macroglobulinemia, unresponsive or relapsed following
 standard of care treatments.

 -  HLA identical (6/6) related donor

 -  For adults: Ability to comprehend the investigational nature of the study and provide
 informed consent. For minors: written informed consent from one parent or guardian.
 Informed oral consent from minors: the process will be explained to the minor on a
 level of complexity appropriate for their age and ability to comprehend.

 EXCLUSION CRITERIA - RECIPIENT:

 -  Estimated probability of surviving less than three months

 -  Major anticipated illness or organ failure incompatible with survival from transplant

 -  Severe psychiatric illness. Mental deficiency sufficiently severe as to make
 compliance with the transplant treatment unlikely and making informed consent
 impossible.

 -  Positive pregnancy test for women of childbearing age.

 -  HIV positive

 -  DLCO adjusted for ventilation and hemoglobin less than 65 percent predicted

 -  Left ventricular ejection fraction less than 40 percent

 -  AST/SGOT greater than 10 times ULN (CTCAE grade IV v3.0)

 -  Bilirubin greater than 5 times ULN (CTCAE grade IV v3.0)

 -  Creatinine greater than 4.5 times ULN (CTCAE grade IV v 3.0)

 -  Prior allogeneic stem cell transplantation.

 INCLUSION CRITERIA - DONOR:

 -  Related donor, HLA identical (6/6) with recipient

 -  Weight greater than or equal to 18 kg

 -  Age greater than or equal to 2 or less than or equal to 80 years old

 -  For adults: Ability to comprehend the investigational nature of the study and provide
 informed consent. For minors: Written informed consent from one parent or guardian and
 informed assent: The process will be explained to the minor on a level of complexity
 appropriate for their age and ability to comprehend.

 EXCLUSION CRITERIA - DONOR:

 -  Pregnant. Lactating donors permitted provide breast milk is discarded during the days
 that G-CSF is given

 -  Unfit to receive filgrastim (G-CSF) and undergo apheresis (abnormal blood counts,
 history of stroke, uncontrolled hypertension)

 -  Sickling hemoglobinopathies including HbSS, HbAS, HbSC

 -  HIV positive donors who are positive for hepatitis B (HBV), hepatitis C (HCV) or human
 T-cell lymphotropic virus (HTLV-I/II) will be used at the discretion of the
 investigator following counseling and approval from the recipient

 -  Severe psychiatric illness. Mental deficiency sufficiently severe as to make
 compliance with the BMT treatment unlikely, and making informed consent impossible