ROLE OF HYPERTHERMIA IN BONE MARROW TRANSPLANTATION

Project: Research project

Description

Autologous bone marrow transplantation is used for many patients including
those with acute myelogenous leukemias and acute lymphoblastic leukemias
who lack appropriate donor marrow. Recurrence of malignancy, however, is a
major problem for patients receiving autologous bone marrow. In this study we will explore and optimize the use of hyperthermia to purge
leukemic cells from bone marrow specimens in vitro. Heat response of human
leukemic cell lines, acute leukemias from patients and normal human bone
marrow progenitors (CFU-GM, BFU-E, CFU-GEMM) will be determined using
clonogenic assays. Single or fractionated doses of heat with or without
chemotherapeutic agents now used in the clinic for in vitro purging of
leukemias will be used. Fractionated doses of heat will be used to take
advantage of the differential rate of decay of thermotolerance of heat
sensitive leukemic cells vs normal bone marrow progenitors. The damage to
the bone marrow stem cells and reconstitution of the purged human bone
marrow will be measured using long-term bone marrow cultures. To ensure the in vivo reconstitution of the purged bone marrow and to
measure the leukemia relapse, a murine leukemia model system will be used.
Both AKR leukemia and murine bone marrow will be treated in vitro with
single or fractionated hyperthermia with or without chemotherapeutic
agents. Furthermore, following purging of leukemia:bone marrow mixtures,
both in vivo reconstitution of the bone marrow and leukemia relapse will be
measured. Finally, we will test the feasibility of using the expression of the
inducible HSP-72 kDa as the predictor of heat response. To determine the
expression of the HSP-72 mRNA, in vitro enzymatic amplification of the HSP-
72 mRNA (PCR) will be performed by the synthetic primers. The inducible
HSP-72 will then be detected by southern blot analysis using synthetic
probes. We will also use 1 & 2 dimensional PAGE to detect the expression
of the HSP-72 kDa protein. The expression of the inducible HSP-72 mRNA and
proteins will then be correlated with the heat response of human leukemia
cells. If there is a positive correlation between the presence of the
inducible HSP-72 kDa and thermal resistance, PCR could be used clinically
to predict the thermal response of leukemic cells before in vitro purging
with heat.
StatusFinished
Effective start/end date7/1/906/30/95

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

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Bone Marrow Transplantation
Fever
Hot Temperature
Bone Marrow
Leukemia
Recurrence
Bone Marrow Cells
Messenger RNA
Bone Marrow Purging
HSP72 Heat-Shock Proteins
Myeloid Progenitor Cells
Polymerase Chain Reaction
Erythroid Precursor Cells
Autologous Transplantation
Southern Blotting
Acute Myeloid Leukemia
Stem Cells
In Vitro Techniques
Tissue Donors
Cell Line

ASJC

  • Medicine(all)