The Common Marmoset as a Target Preclinical Primate Model for Cytokine and Gene Therapy Studies

Abstract

Nonhuman primate models are useful to evaluate the safety and efficacy of new therapeutic modalities, including gene therapy, before the inititation of clinical trials in humans. With the aim of establishing safe and effective approaches to therapeutic gene transfer, we have been focusing on a small New World monkey, the common marmoset, as a target preclinical model. This animal is relatively inexpensive and easy to breed in limited space. First, we characterized marmoset blood and bone marrow progenitor cells (BMPCs) and showed that human cytokines were effective to maintain and stimulate in culture. We then examined their susceptibility to transduction by retroviral vectors. In a mixed culture system containing both marmoset stromal cells and retroviral producer cells, the transduction efficiency into BMPCs and peripheral blood progenitor cells (PBPCs) was 12% to 24%. A series of marmosets then underwent transplantation with autologous PBPCs transduced with a retroviral vector carrying the multidrug resistance 1 gene (MDR1) and were followed for the persistence of these cells in vivo. Proviral DNA was detectable by polymerase chain reaction (PCR) in peripheral blood granulocytes and lymphocytes in the recipients of gene transduced progenitors up to 400 days posttransplantation. To examine the function of the MDR1 gene in vivo, recipient maromsets were challenged with docetaxel, an MDR effluxed drug, yet the overall level of gene transfer attained in vivo (<1% in peripheral blood granulocytes) was not sufficient to prevent the neutropenia induced by docetaxel treatment. Using this model, we safely and easily performed a series of in vivo studies in our small animal center. Our results show that this small nonhuman primate, the common marmoset, is a useful model for the evaluation of gene transfer methods targeting hematopoietic stem cells.

Non-human primate dendritic cells

“…G-CSF, SCF, GM-CSF, Flt3-L, ProGP, and the IL-3/G-CSF chimeric molecule myelopoietin have all been used to mobilize primate stem cells, DC precursors, or in vivo DC, which can potentially be harvested by leukapheresis in larger primate species [15, 47, 49 -53]. G-CSF has also been used in marmoset monkeys to mobilize progenitor cells [54], and this approach can yield sufficient quantities of MoDC and stem cell-derived DCderived from the peripheral blood of this small NWP for in vitro or in vivo studies, without endangering the life of the animal [34].…”

Establishment of Novel Embryonic Stem Cell Lines Derived from the Common Marmoset ( Callithrix jacchus )

“…In addition to these reproductive benefits, we have shown previously that marmosets are suitable laboratory animals for preclinical studies of stem cell therapies, owing to the similarities between the hematopoietic and immune systems of humans and marmosets [4,5]. In 1996, Thomson et al [6] established pluripotent common marmoset cell lines, which are considered powerful tools for understanding the regulatory mechanisms of ESC differentiation both in vitro and in vivo.…”

Tal1/Scl Gene Transduction Using a Lentiviral Vector Stimulates Highly Efficient Hematopoietic Cell Differentiation from Common Marmoset ( Callithrix jacchus ) Embryonic Stem Cells

“…Unlike macaques, marmosets routinely deliver twins or triplets. In addition to these reproductive benefits, similarities between the hematopoietic and immune systems of humans and marmosets make these animals suitable for preclinical studies of stem cell therapies [4,5].…”