Effects of busulfan dose escalation on engraftment of infant rhesus monkey hematopoietic stem cells after gene marking by a lentiviral vector

Christoph Kahl, Alice F. Tarantal, Chang I. Lee, Daniel F. Jimenez, Christopher Choi, Karen Pepper, Denise Petersen, Misty D. Fletcher, Alyssa C. Leapley, Jennifer Fisher, Travis S. Burns, Man Ni Ultsch, Frederick J. Dorey, Donald B. Kohn

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Objective. Nonmyeloablative cytoreduction is used in clinical hematopoietic stem cell gene therapy trials to increase engraftment of gene-modified cells. We utilized an infant rhesus monkey model to identify an optimal dosage of busulfan that results in efficient long-term gene marking with minimal toxicities. Methods. Bone marrow (BM) was harvested, followed by a single 2-hour intravenous infusion of busulfan at escalating dosages of 0 to 160 mg/m 2. CD34 + cells were immunoselected from BM, transduced overnight with a simian immunodeficiency virus-based lentiviral vector carrying a nonexpressed marker gene, and injected intravenously 48 hours post-busulfan administration. Pharmacokinetics were assessed, as well as adverse effects and peripheral blood and BM gene marking. Results. Increasing dosages of busulfan resulted in increased area-under-the-curve (AUC) with some variability at each dosage level, suggesting interindividual variation in clearance. Blood chemistries were normal and no adverse effects were observed as a result of busulfan infusion. At 120 and 160 mg/m 2, transient neutropenia and thrombocytopenia were noted but not lymphopenia. Over the 6 months of study posttransplantation, a busulfan dosage-related increase in gene marking was observed ranging from undetectable (no busulfan) up to 0.1% gene-containing cells in animals achieving the highest busulfan AUC. This corresponds to a more than 100-fold increase in gene marking over the busulfan dosage range studied. Conclusions. These data indicate that increased gene marking of hematopoietic stem cells can be achieved by escalating busulfan dosages from 40 to 160 mg/m 2 without significant toxicity in infant nonhuman primates.

Original languageEnglish (US)
Pages (from-to)369-381
Number of pages13
JournalExperimental Hematology
Volume34
Issue number3
DOIs
StatePublished - Mar 2006
Externally publishedYes

Fingerprint

Busulfan
Hematopoietic Stem Cells
Macaca mulatta
Genes
Bone Marrow
Area Under Curve
Simian Immunodeficiency Virus
Lymphopenia
Cell- and Tissue-Based Therapy
Neutropenia
Intravenous Infusions
Thrombocytopenia
Genetic Therapy
Primates
Pharmacokinetics

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

Effects of busulfan dose escalation on engraftment of infant rhesus monkey hematopoietic stem cells after gene marking by a lentiviral vector. / Kahl, Christoph; Tarantal, Alice F.; Lee, Chang I.; Jimenez, Daniel F.; Choi, Christopher; Pepper, Karen; Petersen, Denise; Fletcher, Misty D.; Leapley, Alyssa C.; Fisher, Jennifer; Burns, Travis S.; Ultsch, Man Ni; Dorey, Frederick J.; Kohn, Donald B.

In: Experimental Hematology, Vol. 34, No. 3, 03.2006, p. 369-381.

Research output: Contribution to journalArticle

Kahl, C, Tarantal, AF, Lee, CI, Jimenez, DF, Choi, C, Pepper, K, Petersen, D, Fletcher, MD, Leapley, AC, Fisher, J, Burns, TS, Ultsch, MN, Dorey, FJ & Kohn, DB 2006, 'Effects of busulfan dose escalation on engraftment of infant rhesus monkey hematopoietic stem cells after gene marking by a lentiviral vector', Experimental Hematology, vol. 34, no. 3, pp. 369-381. https://doi.org/10.1016/j.exphem.2005.12.005
Kahl, Christoph ; Tarantal, Alice F. ; Lee, Chang I. ; Jimenez, Daniel F. ; Choi, Christopher ; Pepper, Karen ; Petersen, Denise ; Fletcher, Misty D. ; Leapley, Alyssa C. ; Fisher, Jennifer ; Burns, Travis S. ; Ultsch, Man Ni ; Dorey, Frederick J. ; Kohn, Donald B. / Effects of busulfan dose escalation on engraftment of infant rhesus monkey hematopoietic stem cells after gene marking by a lentiviral vector. In: Experimental Hematology. 2006 ; Vol. 34, No. 3. pp. 369-381.
@article{a671960c73a34a9f9fea16f63bc7aeff,
title = "Effects of busulfan dose escalation on engraftment of infant rhesus monkey hematopoietic stem cells after gene marking by a lentiviral vector",
abstract = "Objective. Nonmyeloablative cytoreduction is used in clinical hematopoietic stem cell gene therapy trials to increase engraftment of gene-modified cells. We utilized an infant rhesus monkey model to identify an optimal dosage of busulfan that results in efficient long-term gene marking with minimal toxicities. Methods. Bone marrow (BM) was harvested, followed by a single 2-hour intravenous infusion of busulfan at escalating dosages of 0 to 160 mg/m 2. CD34 + cells were immunoselected from BM, transduced overnight with a simian immunodeficiency virus-based lentiviral vector carrying a nonexpressed marker gene, and injected intravenously 48 hours post-busulfan administration. Pharmacokinetics were assessed, as well as adverse effects and peripheral blood and BM gene marking. Results. Increasing dosages of busulfan resulted in increased area-under-the-curve (AUC) with some variability at each dosage level, suggesting interindividual variation in clearance. Blood chemistries were normal and no adverse effects were observed as a result of busulfan infusion. At 120 and 160 mg/m 2, transient neutropenia and thrombocytopenia were noted but not lymphopenia. Over the 6 months of study posttransplantation, a busulfan dosage-related increase in gene marking was observed ranging from undetectable (no busulfan) up to 0.1{\%} gene-containing cells in animals achieving the highest busulfan AUC. This corresponds to a more than 100-fold increase in gene marking over the busulfan dosage range studied. Conclusions. These data indicate that increased gene marking of hematopoietic stem cells can be achieved by escalating busulfan dosages from 40 to 160 mg/m 2 without significant toxicity in infant nonhuman primates.",
author = "Christoph Kahl and Tarantal, {Alice F.} and Lee, {Chang I.} and Jimenez, {Daniel F.} and Christopher Choi and Karen Pepper and Denise Petersen and Fletcher, {Misty D.} and Leapley, {Alyssa C.} and Jennifer Fisher and Burns, {Travis S.} and Ultsch, {Man Ni} and Dorey, {Frederick J.} and Kohn, {Donald B.}",
year = "2006",
month = "3",
doi = "10.1016/j.exphem.2005.12.005",
language = "English (US)",
volume = "34",
pages = "369--381",
journal = "Experimental Hematology",
issn = "0301-472X",
publisher = "Elsevier Inc.",
number = "3",

}

TY - JOUR

T1 - Effects of busulfan dose escalation on engraftment of infant rhesus monkey hematopoietic stem cells after gene marking by a lentiviral vector

AU - Kahl, Christoph

AU - Tarantal, Alice F.

AU - Lee, Chang I.

AU - Jimenez, Daniel F.

AU - Choi, Christopher

AU - Pepper, Karen

AU - Petersen, Denise

AU - Fletcher, Misty D.

AU - Leapley, Alyssa C.

AU - Fisher, Jennifer

AU - Burns, Travis S.

AU - Ultsch, Man Ni

AU - Dorey, Frederick J.

AU - Kohn, Donald B.

PY - 2006/3

Y1 - 2006/3

N2 - Objective. Nonmyeloablative cytoreduction is used in clinical hematopoietic stem cell gene therapy trials to increase engraftment of gene-modified cells. We utilized an infant rhesus monkey model to identify an optimal dosage of busulfan that results in efficient long-term gene marking with minimal toxicities. Methods. Bone marrow (BM) was harvested, followed by a single 2-hour intravenous infusion of busulfan at escalating dosages of 0 to 160 mg/m 2. CD34 + cells were immunoselected from BM, transduced overnight with a simian immunodeficiency virus-based lentiviral vector carrying a nonexpressed marker gene, and injected intravenously 48 hours post-busulfan administration. Pharmacokinetics were assessed, as well as adverse effects and peripheral blood and BM gene marking. Results. Increasing dosages of busulfan resulted in increased area-under-the-curve (AUC) with some variability at each dosage level, suggesting interindividual variation in clearance. Blood chemistries were normal and no adverse effects were observed as a result of busulfan infusion. At 120 and 160 mg/m 2, transient neutropenia and thrombocytopenia were noted but not lymphopenia. Over the 6 months of study posttransplantation, a busulfan dosage-related increase in gene marking was observed ranging from undetectable (no busulfan) up to 0.1% gene-containing cells in animals achieving the highest busulfan AUC. This corresponds to a more than 100-fold increase in gene marking over the busulfan dosage range studied. Conclusions. These data indicate that increased gene marking of hematopoietic stem cells can be achieved by escalating busulfan dosages from 40 to 160 mg/m 2 without significant toxicity in infant nonhuman primates.

AB - Objective. Nonmyeloablative cytoreduction is used in clinical hematopoietic stem cell gene therapy trials to increase engraftment of gene-modified cells. We utilized an infant rhesus monkey model to identify an optimal dosage of busulfan that results in efficient long-term gene marking with minimal toxicities. Methods. Bone marrow (BM) was harvested, followed by a single 2-hour intravenous infusion of busulfan at escalating dosages of 0 to 160 mg/m 2. CD34 + cells were immunoselected from BM, transduced overnight with a simian immunodeficiency virus-based lentiviral vector carrying a nonexpressed marker gene, and injected intravenously 48 hours post-busulfan administration. Pharmacokinetics were assessed, as well as adverse effects and peripheral blood and BM gene marking. Results. Increasing dosages of busulfan resulted in increased area-under-the-curve (AUC) with some variability at each dosage level, suggesting interindividual variation in clearance. Blood chemistries were normal and no adverse effects were observed as a result of busulfan infusion. At 120 and 160 mg/m 2, transient neutropenia and thrombocytopenia were noted but not lymphopenia. Over the 6 months of study posttransplantation, a busulfan dosage-related increase in gene marking was observed ranging from undetectable (no busulfan) up to 0.1% gene-containing cells in animals achieving the highest busulfan AUC. This corresponds to a more than 100-fold increase in gene marking over the busulfan dosage range studied. Conclusions. These data indicate that increased gene marking of hematopoietic stem cells can be achieved by escalating busulfan dosages from 40 to 160 mg/m 2 without significant toxicity in infant nonhuman primates.

UR - http://www.scopus.com/inward/record.url?scp=33644923202&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33644923202&partnerID=8YFLogxK

U2 - 10.1016/j.exphem.2005.12.005

DO - 10.1016/j.exphem.2005.12.005

M3 - Article

C2 - 16543071

AN - SCOPUS:33644923202

VL - 34

SP - 369

EP - 381

JO - Experimental Hematology

JF - Experimental Hematology

SN - 0301-472X

IS - 3

ER -