Journal: bioRxiv

Article Title: JARID2 Inhibition Reprograms Human Hematopoietic Progenitor Cells To Enhance Bone Marrow Transplantation

doi: 10.1101/2025.09.03.672211

Figure Lengend Snippet: (a) Longitudinal analysis of human CD45+ chimerism in peripheral blood (PB) of NSG mice transplanted with cord blood CD34+ cells targeted with guide RNAs for AAVS1 (n=4), JARID2 #1 (n=5) or JARID2 #2 (n=5). (b) Longitudinal analysis of modified alleles from CRISPR edits in human CD45+ PB cells in transplanted NSG mice. (c) Lineage composition of human CD45+ PB cells one-year post-transplant (B-cells = CD19+, T-cells = CD3+, myeloid cells = CD33+). (d) Complete blood count (CBC) analysis of white blood cells (WBC), red blood cells (RBC), hemoglobin (Hb), hematocrit (HCT) and platelets (PLT) of recipient mice one-year post-transplant. Dashed lines represent normal range. (e) Representative histology of PB smear, bone marrow (BM) cytospins, and spleen (SPL) harvested one-year after transplantation of CRISPR-edited human CD34+ cells into NSG mice. Scale bars - 10µm (PB) and 25µm (BM and SPL).

Article Snippet: The following antibodies were used for profiling chromatin-bound factors: anti-CTCF (Cell Signaling Technology, #2899S), EZH2 (Abcam, #ab3748; CST, #5246S), H2AK119ub1 (CST, #8240S), histone H3 (CST, #4499S), H3K27ac (CST, #8173S), H3K27me3 (CST, #9733S), H3K4me3 (CST, #9751S; EpiCypher, #13-0060), and JARID2 (Asp1114) (CST, #13594S).

Techniques: Modification, CRISPR, Transplantation Assay

Journal: bioRxiv

Article Title: JARID2 Inhibition Reprograms Human Hematopoietic Progenitor Cells To Enhance Bone Marrow Transplantation

doi: 10.1101/2025.09.03.672211

Figure Lengend Snippet: (a) Peripheral blood (PB) engraftment of cord blood cells transduced with indicated constitutive lentiviral shRNAs. (b) Lineage composition of GFP+ PB cells 16-weeks post-transplant. (c) Engraftment of GFP+ human cells in the BM of recipient mice 20-weeks post-transplant. (d) Quantification of GFP+ human HSPC populations in the BM of recipient mice 20-weeks post-transplant (sh Ren n=16, sh JARID2 #1 n=14, sh JARID2 #3 n=17). (e) PB engraftment of cord blood cells transduced with indicated constitutive lentiviral shRNAs in secondary recipients. (f) Lineage composition of GFP+ PB cells 16-weeks post-secondary transplant. (g) Engraftment of GFP+ human cells in the BM of secondary recipient mice 20-weeks post-transplant. (h) Quantification of GFP+ human HSPC populations in the BM of recipient mice 20-weeks post-transplant (sh Ren n=8, sh JARID2 #1 n=6, sh JARID2 #3 n=6).

Article Snippet: The following antibodies were used for profiling chromatin-bound factors: anti-CTCF (Cell Signaling Technology, #2899S), EZH2 (Abcam, #ab3748; CST, #5246S), H2AK119ub1 (CST, #8240S), histone H3 (CST, #4499S), H3K27ac (CST, #8173S), H3K27me3 (CST, #9733S), H3K4me3 (CST, #9751S; EpiCypher, #13-0060), and JARID2 (Asp1114) (CST, #13594S).

Techniques: Transduction

Journal: bioRxiv

Article Title: JARID2 Inhibition Reprograms Human Hematopoietic Progenitor Cells To Enhance Bone Marrow Transplantation

doi: 10.1101/2025.09.03.672211

Figure Lengend Snippet: (a) tSNE plots of lineage distribution of human CD45+ cells in BM of recipient mice 20-weeks post-transplant by flow cytometry. (b) Frequency of maturing myeloid cells in NSGS mice transplanted with cord blood cells transduced with indicated constitutive lentiviral shRNAs 20-weeks post-transplant. (c) JARID2 expression across indicated cell populations showing knockdown efficiency. (d) UMAP of scRNA-seq data for hCD34+ cells isolated 20-weeks post-transplantation. Cell populations are identified by marker genes. (e) Numbers of cells of indicated genotypes per cluster based on Azimuth cell type annotations.

Article Snippet: The following antibodies were used for profiling chromatin-bound factors: anti-CTCF (Cell Signaling Technology, #2899S), EZH2 (Abcam, #ab3748; CST, #5246S), H2AK119ub1 (CST, #8240S), histone H3 (CST, #4499S), H3K27ac (CST, #8173S), H3K27me3 (CST, #9733S), H3K4me3 (CST, #9751S; EpiCypher, #13-0060), and JARID2 (Asp1114) (CST, #13594S).

Techniques: Flow Cytometry, Transduction, Expressing, Knockdown, Isolation, Transplantation Assay, Marker

Journal: bioRxiv

Article Title: JARID2 Inhibition Reprograms Human Hematopoietic Progenitor Cells To Enhance Bone Marrow Transplantation

doi: 10.1101/2025.09.03.672211

Figure Lengend Snippet: (a) Peripheral blood (PB) engraftment of cord blood cells transduced with indicated inducible lentiviral shRNAs induced for eight-days ex vivo prior to transplant. (b) Lineage composition of hD45+ PB cells 16-weeks post-transplant. (c) Engraftment of hCD45+ cells in the BM of recipient mice 20-weeks post-transplant. (d) Quantification of hCD45+ HSPC populations in the BM of recipient mice 20-weeks post-transplant (sh Ren n=9, sh JARID2 #1 n=6, sh JARID2 #3 n=8, sh EZH2 n=7). (e) PB engraftment of cord blood cells transduced with indicated inducible shRNAs induced in vivo at 14-weeks post-transplant (normalized to sh Ren ). (f) Engraftment of BFP+ hCD45+ cells in the BM of recipient mice (shRNAs for JARID2 and EZH2 are combined for statistical comparison). (g) Quantification of BFP+ hCD45+ HSPC populations in the BM of recipient mice (sh Ren n=3, sh JARID2 #1+3 n=9, sh EZH2 #1+2 n=8).

Article Snippet: The following antibodies were used for profiling chromatin-bound factors: anti-CTCF (Cell Signaling Technology, #2899S), EZH2 (Abcam, #ab3748; CST, #5246S), H2AK119ub1 (CST, #8240S), histone H3 (CST, #4499S), H3K27ac (CST, #8173S), H3K27me3 (CST, #9733S), H3K4me3 (CST, #9751S; EpiCypher, #13-0060), and JARID2 (Asp1114) (CST, #13594S).

Techniques: Transduction, Ex Vivo, In Vivo, Comparison

Journal: bioRxiv

Article Title: JARID2 Inhibition Reprograms Human Hematopoietic Progenitor Cells To Enhance Bone Marrow Transplantation

doi: 10.1101/2025.09.03.672211

Figure Lengend Snippet: (a) UMAP of scRNA-seq data for hCD34+ cells sorted from NSG mice transplanted with cord blood cells transduced with constitutive lentiviral shRNAs 20-weeks post-transplant. Cell populations are identified by marker genes. sh Ren , sh JARID2 #1 and sh JARID2 #3 samples are combined, the HSC population is highlighted. (b) Dot plot highlighting differential expression of a curated HSC signature gene set (Aguadé-Gorgorió et al., 2024, Nature) amongst shRNA genotypes within the HSC population. (c) HSC module scores calculated using the Seurat package based on the curated HSC signature gene list. (d) Serial replating showing CFU efficiency of indicated HSPC populations isolated from BM of recipient mice 20-weeks post-transplant. (e) Volcano plot showing differentially expressed genes between sh JARID2 and sh Ren cells within the HSC population. (f) Representative flow cytometry plots showing the proportion of MHCII-high (red boxes) HSCs from each genotype from the BM of recipient mice 20-weeks post-transplant. (g) Serial replating showing CFU efficiency of indicated HSPC genotypes separated by MHCII levels isolated from BM of recipient mice 20-weeks post-transplant. (h) Cell cycle analysis by flow cytometry showing proportion of quiescent (G 0 ) HSCs of each genotype from the BM of recipient mice 20-weeks post-transplant.

Article Snippet: The following antibodies were used for profiling chromatin-bound factors: anti-CTCF (Cell Signaling Technology, #2899S), EZH2 (Abcam, #ab3748; CST, #5246S), H2AK119ub1 (CST, #8240S), histone H3 (CST, #4499S), H3K27ac (CST, #8173S), H3K27me3 (CST, #9733S), H3K4me3 (CST, #9751S; EpiCypher, #13-0060), and JARID2 (Asp1114) (CST, #13594S).

Techniques: Transduction, Marker, Quantitative Proteomics, shRNA, Isolation, Flow Cytometry, Cell Cycle Assay