Figure 6.

Stable Promoter Activity and Polyclonal Hematopoiesis Using the Non-insulated MND LV in a Long-Term NHP Gene Therapy Model

(A) Schematic diagram of SIN-LV used in competitive repopulation study performed with NHP autologous BM, GFP, yellow fluorescent protein (YFP), and the 1.6-kb proximal human WAS promoter (WS1.6).⁵⁷ (B) Percentage of cells expressing the indicated fluorescent protein in peripheral blood mononuclear cells (PBMCs) over time after transplant. (C) Representative flow plots showing marking (bottom row) within specific PB subsets (top row) obtained 326 days post-transplant. (D) The MFI for each sub-population in this sample is shown. (E) Frequency of MSG-PCR clones identified in sorted GFP⁺ or YFP⁺ PB cells at 502, 952, or 1,153 days after transplant. The total number of clones sequenced is indicated at the top of each bar in the chart. Thickness of bar segments indicates the frequency of a particular clone; the color given to each clone is consistent for comparison between time points. (F) Circos plot mapping RIS positions identified in GFP⁺ (green histogram) and YFP⁺ (yellow histogram) PB cells (obtained between 502 and 1,153 days after gene therapy) to the Rhesus genome (outer circle showing chromosome numbers). The Rhesus genome was divided into 1-Mbp bins; the heights of the histograms indicate the number of integrations found in a 1-Mbp bin.