Prolonged multilineage clonal hematopoiesis in a rhesus recipient of CD34 positive cells marked with a RD114 pseudotyped oncoretroviral vector

Acute myeloid leukemia is associated with retroviral gene transfer to hematopoietic progenitor cells in a rhesus macaque

“…These 2 insertions were also identified by LAM-PCR analysis on DNA from individual blasts obtained via single-cell laser capture on kidney sections. PCR analysis using primers specific for the insertion into BCL2-A1 on chromosome 15q25.1 and the insertion into chromosome 9q31.1 revealed a similar pattern of high level contribution from the dominant clone in the blood during the first year after transplantation as previously reported, 4 which then became undetectable, before dominating again in the blood at the time of myeloid sarcoma manifestation (Figure S3A-B). TaqMan reverse transcriptase (RT)-PCR was used to assess BCL2-A1 mRNA expression from bulk tumor RNA revealing levels comparable to granulocytes, where BCL2-A1 is highly expressed, and about 70 times higher than in the kidney tissue of an untreated control animal.…”

“…The animal remained clinically well and had normal blood counts throughout. 4 It received one cycle of chemotherapy with transient in vivo selection of drug resistant cells 2 years after transplantation. 5 At 5 years after transplantation, 96E113 developed a left renal mass impinging on intestines and pancreas.…”

“…7 Mature blood cells and enriched blasts (Ͼ 95%) from kidney and liver infiltrates have been analyzed by linear amplification mediated (LAM)-PCR and inverse PCR, topoisomerase I-thymidine-adenine (TOPO TA) cloning and subsequent sequencing as described. 4,8 The insertion sites were mapped against the current build of the human genome as well as the preliminary rhesus draft genome. 9,10 In blood and tumor cells, the same vector insertions characterizing the dominant clone during the first year after transplantation were again identified.…”

Improved transduction of human sheep repopulating cells by retrovirus vectors pseudotyped with feline leukemia virus type C or RD114 envelopes

“…[59][60][61][62][63][64] Our observations about superior transduction with RD114-pseudotyped retrovirus particles are consistent with previous data in canine, nonhuman primate, and NOD-SCID mouse xenograft model systems. [40][41][42][43][44][45] Our data are the first demonstration of transduction of primary hematopoietic cells with recombinant FeLV-C retrovirus particles. We conclude that FeLV-C-and RD114-pseudotyped virus particles transduce human stem and progenitor cells more efficiently than GALV-pseudotyped retrovirus particles, which may allow gene therapy to be extended to hematopoietic diseases other than SCID.…”

“…Recombinant retrovirus vectors with envelopes derived from the RD114 virus have been used successfully to transduce primitive human, canine, and primate hematopoietic cells, often at high levels. [40][41][42][43][44][45] Feline leukemia virus type C (FeLV-C) causes redcell aplasia in cats. The FeLV-C envelope recognizes a heme transporter protein (FeLV-C receptor [FLVCR]).…”