Mobilization and collection of peripheral blood CD34+ cells from patients with Fanconi anemia

Fanconi anemia: a model disease for studies on human genetics and advanced therapeutics

“…To overcome these limitations, FA gene therapy clinical trials are in progress [85]. Difficulties in collecting sufficient blood progenitor cells from patients with FA and inefficient transduction protocols with first-generation retroviral vectors led to unsuccessful initial clinical trials [86][87][88].…”

Rapid 1-hour transduction of whole bone marrow leads to long-term repopulation of murine recipients with lentivirus-modified hematopoietic stem cells

“…[1][2][3][4] Further, extended ex vivo manipulation can be a critical obstacle in candidate applications with exhausted autologous stem cell pools, such as Fanconi anemia (FA), providing the added rationale to develop rapid transduction protocols. 5,6 Some of these limitations can be overcome by human immunodeficiency virus (HIV)-derived lentivirus vectors with their ability to transduce nondividing cells, thereby permitting shortened ex vivo culture durations while maintaining gene transfer to long-term repopulating cells. 7 We have previously reported long-term gene transfer rates of 12-40% after VSV-G/ lentivirus vector transduction of murine whole bone marrow cells after 12-h prestimulation in murine stem cell factor (mSCF) and a single subsequent 12-h vector exposure.…”

AMD3100 synergizes with G-CSF to mobilize repopulating stem cells in Fanconi anemia knockout mice

“…As expected, the repopulating ability of the BM from Fanca −/− and Fancc −/− mice was only 35-40% of syngeneic WT controls (Table I). Strikingly, there was a much greater defect in the repopulating activity of Fanca −/− and Fancc −/− PB cells following mobilization with G-CSF alone (15-30% of WT controls) analogous to the paucity of CD34 + phenotypic cells mobilized using G-CSF only in FA patients [32] and the reduced numbers of lineage restricted and multipotent progenitors. Though stem cells mobilized with AMD3100 alone resulted in low chimerism in all experimental groups (Figure 3), addition of G-CSF to AMD3100 resulted in a greater than additive increase in the chimerism and repopulating ability of Fanca −/− and Fancc −/− HSC ( Figure 3, Table I).…”

“…Genetic correction and transplantation of autologous HSC is a potential therapy; however, mobilization of adequate HSC in FA patients is a key prerequisite for non-invasive collection. Previous trials in FA patients using extended G-CSF administration mobilized a limited number of CD34 + cells [32]. It was unclear whether the relative inability of G-CSF to mobilize CD34 + cells in FA patients is a consequence of BM hypoplasia or a defect in stem cell/stromal cell interactions.…”

“…Granulocyte colonystimulating factor (G-CSF) is an FDA approved therapy utilized for mobilization of hematopoietic stem (HSC)/progenitor cells (HPC) for autologous transplantation in a variety of clinical settings [31]. However, patients with FA show a markedly decreased response to G-CSF [32]. The inability of G-CSF to adequately mobilize HSC/HPC in FA patients has prompted the search for alternative methods.…”