Transplantation of Umbilical Cord Blood Following Chemotherapy for Blood Cancers

The success of BMT as a curative option for patients with malignancies is frequently limited
by the inability to identify an appropriate donor in time for transplantation.

Transplantations utilizing umbilical cord blood stem cells are increasingly successful. Data
suggest that there are fewer and less developed T-cells in cord products when compared to
bone marrow and so are likely to produce less graft-vs.-host disease (GVHD) even in the
mismatched setting. In addition, the grafts are easily available second to the ease of
collection as compared with other sources such as bone marrow and peripheral blood stem cells
(PBSCs). The ability to build cord blood repositories containing samples with HLA types from
minorities has and will continue to increase the likelihood of finding suitable products for
under represented minorities such as African-Americans, Hispanic populations and mixed ethnic
populations.

Unfortunately, limitations remain, secondary to the time to engraftment when the cell dose is
less than optimal, resulting in delayed or failed engraftment. In addition, increased cell
dose appears to be associated with disease free survival. Therefore, this modality of
transplant is generally limited to smaller patients. This protocol evaluates the infusion and
engraftment of two cord blood products - one that has been expanded ex vivo and one that has
not been expanded following myeloablative chemotherapy for the treatment of hematological
malignancies.

Primary Objective:

-To evaluate the safety of transplantation of two cord products including an expanded unit
including infusional toxicities and potential immunologic competition.

Secondary Objective:

-To determine if the use of two cord products results in an improvement in neutrophil
engraftment (ANC>500) in subjects as demonstrated by engraftment <=21 days.

Tertiary Objectives:

Patients will be compared with patients participating in other trials at our institution and
to those who receive therapy as per standard of care.

- To evaluate the rate, extent, and durability of hematopoietic reconstitution and in
particular to determine the relative contribution of each cord unit in early engraftment
and long-term engraftment

- To estimate the rate and severity of graft-vs-host disease

- To estimate the rates of infectious complications

- To obtain preliminary data regarding disease-free and overall survival

- To collect samples for future studies of immune reconstitution and for future studies in
the laboratory regarding disease and the immune system

We will be able to track the fate of both the unmanipulated and expanded CB progenitors by
evaluating differences in the cord products and the recipient by evaluation of sex mismatch,
HLA type and/or restriction length polymorphisms (RFLPs). If the expanded CB cells are
detected in the patients long-term, this will give us confidence to use expanded CB as the
sole hematopoietic progenitor cell support, in future studies.

The major risk is non-engraftment of either one or both of the cord blood units.
Non-engraftment of one of the units may lead to prolonged cytopenia and a marked increased
risk for infection. Failure of either unit to engraft as defined as failure to detect cells
from either cord blood product at Day +60 is likely to result in the death of the subject.
There is the possibility of failure of long-term engraftment from both cords. This would be
fatal unless an alternative donor option was identified or the unlikely event of autologous
reconstitution of bone marrow. There is a possibility of an immune competition of both cord
blood products . This could result in the loss of the cord product that would have been
responsible for long term engraftment. There is a possibility of the expanded cord product to
fail release testing. In this event, only the unexpanded cord would be infused and would
likely result in delayed engraftment.

As with all cellular products there are risks associated with the infusion including but not
limited to anaphylaxis, transfusion reaction, hemolysis, side-effects of the DMSO that the
product is stored in (bradycardia, hypothermia, neurologic changes). The expanded product
will be washed however there remains a small chance that a small amount of growth factor
remains. The risk associated with these will be allergic.

Busulfan toxicity

1. Nausea and vomiting

2. Myeloablation

3. Pulmonary complications

4. Seizures

5. Other toxic effects which may be produced by Busulfan erythematous skin rash,
hyperpigmentation, hepatic dysfunction, amenorrhea, testicular atrophy, gynecomastia,
myasthenia symptoms, cataract and atrophic bronchitis associated with cytologic
dysplasia.

Cyclophosphamide Toxicity

1. Leukopenia, anemia

2. Alopecia

3. Nausea, vomiting, increased AST, ALT, mucositis, diarrhea

4. Headache, dizziness

5. Cardiac necrosis rarely with high dose cyclophosphamide

6. Hemorrhagic cystitis, SIADH

7. Teratogenic, may cause secondary neoplasms, anaphylaxis (rare)

8. Fluid retention.

9. Cardiomyopathy.

10. Hemorrhagic Cystitis.

At the doses used for uroprotection mesna is virtually non-toxic. However, adverse effects
which may be attributable to mesna include nausea and vomiting, diarrhea, abdominal pain,
altered taste, rash, urticaria, headache, joint or limb pain, hypotension and fatigue.

Total Body Irradiation Toxicity:

1. Nausea and Vomiting

2. Alopecia

3. Parotitis and Pancreatitis

4. Diarrhea

5. Fever

6. Erythema

7. Hyperpigmentation

8. Mucositis

9. Late Effects - cataract formation, growth retardation, carcinogenesis and the
probability of sterilization.

Inclusion Criteria:

- Patient must have two cord units available. Units must be minimally matched to the
subject at 4/6 antigens (HLA Class I (A or B) and Class II (DRB1) - units must have at
lease one HLA DRB1 matched allele) and at least one unit must contain a minimum of 1.0
x 107 Total Nucleated Cells /Kg but neither unit may have > 5 x 107Total Nucleated
Cells /Kg. (The feasibility of using particular units will be discussed with the
Principal Investigators)

- Disease status precludes waiting to identify a suitably HLA matched unrelated donor

- Patients must have a diagnosis of one of the following:

- AML

- refractory AML

- Secondary AML

- ALL in CR2 with high-risk features such as short CR1 and/or high-risk
cytogenetics

- ALL in CR1 following initial induction failure

- Acute mixed lineage leukemia

- CML beyond chronic phase 1.

- Lymphoma (Hodgkins or Non-Hodgkins) ineligible for Autologous-BMT

- Myelodysplastic Syndrome

- Able to provide informed consent or parent/guardian able to provide informed consent.

Exclusion Criteria:

- Consenting 5/6 or 6/6 HLA matched related donor available

- Single cord blood product with cell count >5 x10E7 Total Nucleated Cells/kg

- Poor Performance Status: ECOG performance status >= 2 (Karnofsky or Lansky Play
performance<70).

- Poor Cardiac Function (obtained within 3 weeks of the start of transplant): Left
ventricular ejection fraction <= 45% as determined by MUGA or ECHO. For pediatric
patients LVEF < 45 % or a Shortening Fraction below normal limits for age.

- Poor Pulmonary Function (obtained within 3 weeks of the start of transplant):

- FEV1 and FVC <50% of predicted for patients who have not received thoracic or
mantle irradiation.

- For patients who have received thoracic or mantle irradiation, FEV1 and FVC <=
75% of predicted or DLCO <= 50% of predicted

- For children unable to perform PFTs second to developmental stage, Pulse oximetry
<= 85% on RA

- Poor Liver Function (obtained within 1 week of the start of transplant): Bilirubin >=
2.0 mg/dl. (with the exception of patients whose hyperbilirubinemia is the result of
Gilbert's disease)

- Poor Renal Function (obtained within 3 weeks of the start of transplant): Corrected
CrCl < 60 mg/min. CrCl will be estimated by the Schwartz formula. A measured CrCl or a
GFR may be substituted to determine the subject's CrCl

- HIV Infection: Patients who are HIV positive. (The role of allogenic transplant in
HIV+ individuals has not been studied)

- Pregnancy: Patients who are pregnant. (The chemotherapeutic agents used in bone marrow
transplant are teratogenic)

- Uncontrolled viral, bacterial or fungal infections

- Patients with symptoms consistent with RSV, influenza A, B or parainfluenza at the
time of enrollment on this study will be assayed for the above viruses and if positive
are not eligible for the trial until are no longer symptomatic (patients may have
continued assay positivity for a period of time post resolution of symptoms second to
the nature of the assay)

- Presence of concomitant medication or incident condition that would create an
unreasonable risk for the subject to participate in this study as determined by the
investigators (Primary or co-investigators).

- Patients with known intolerance to or contraindication for any agent that will be used
in the subject's proposed myeloablative or required supportive care regimen.