Journal: Molecular Therapy Oncology

Article Title: Co-editing of NKG2A and FAS increases long-term cytotoxic capacity and persistence of CAR NK cells

doi: 10.1016/j.omton.2026.201126

Figure Lengend Snippet: NKG2A-edited CAR NK cells eliminate CD19-negative target cells resistant to conventional CAR NK cells (A) Specific lysis of 721.45 CD19 KO cells by engineered NK cells (left) or CAR NK cells (right). Cells were co-cultured for 4–5 h at an effector to target ratio of 5:1. (B) Schematic of repetitive stimulation assay. 0.1 × 10 6 NK or CAR NK cells were seeded at day 0, and 0.1 × 10 6 CD19-positive 721.45 cells (target cells) were added every 2–3 days for 14 days. IL-2 was added 2×/week until day 12. Target cells added on day 7 were labeled with PKH26 and on day 14 with PKH67. Data shown in (C–H) were generated using this setup. (C) Expression of CD19 (histograms) and relative MFI of CD19 on total target cells (live CD56 − ) and representative flow cytometry plots on day 14 after co-culture with NK (top) or CAR NK (bottom) cells. Relative MFI is calculated by dividing by MFI of CD19 on target cells cultured alone. (D) Representative expression of PKH26 (target cells added day 7) and PKH67 (target cells added day 14) on total target cells (live CD56 − ) after co-culture with NK or CAR NK cells for 14 days. (E and F) Frequency of remaining live PKH26 + target cells (added on day 7) on day 14 after co-culture with NK cells (E) or CAR NK cells (F). Target cells alone (721.45 only) were not included in the statistical analysis. (G and H) Frequency of remaining live PKH67 + target cells (added on day 14 and co-cultured for 4–5 h before readout) after co-culture with NK cells (G) or CAR NK cells (H). Target cells alone (721.45 only) were not included in the statistical analysis. (A–H) n = 6–9 (individual donors). One-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: EBV-transformed LCLs from allogeneic human fetal liver tissue were generated by isolating CD19 + cells by magnetic-activated cell sorting with CD19 beads (Miltenyi, Cat. 130-050-301) and then infected with EBV B95-8, produced as previously described at a multiplicity of infection (MOI) of 0.05–0.15.

Techniques: Lysis, Cell Culture, Labeling, Generated, Expressing, Flow Cytometry, Co-Culture Assay

Journal: Molecular Therapy Oncology

Article Title: Co-editing of NKG2A and FAS increases long-term cytotoxic capacity and persistence of CAR NK cells

doi: 10.1016/j.omton.2026.201126

Figure Lengend Snippet: Increased cytotoxicity of NKG2A-edited CAR NK cells against CD19-negative BCP-ALL PDX (A) CD19 and (B) HLA-E expression of BCP-ALL PDX by flow cytometry. (C) Specific lysis of PDX1 and PDX2 after co-culture with control NK cells (NK mock) or CAR NK cells (CAR mock). Co-culture at E:T ratio 5:1 for 4 h. (D) Specific lysis of PDX1 after 4 h co-culture at 1:1 E:T ratio. (E) Data from (D) shown as relative increase in killing of engineered CAR NK cells compared with controls (CAR mock). (F–H) Specific lysis of PDX2 after 20 h co-culture at 5:1 E:T ratio with NK (F) or CAR NK (G) cells. (H) Data from (G) shown as relative increase in killing of engineered CAR NK cells compared with controls (CAR mock). (C) n = 3 (individual donors), paired t test. (D–G) n = 3 (individual donors), repeated measures one-way ANOVA. Engineered NK cells shown in this figure were generated from bulk NK cells (not NKG2A + NK cells) as the starting population. ∗ p < 0.05.

Article Snippet: EBV-transformed LCLs from allogeneic human fetal liver tissue were generated by isolating CD19 + cells by magnetic-activated cell sorting with CD19 beads (Miltenyi, Cat. 130-050-301) and then infected with EBV B95-8, produced as previously described at a multiplicity of infection (MOI) of 0.05–0.15.

Techniques: Expressing, Flow Cytometry, Lysis, Co-Culture Assay, Control, Generated

Journal: bioRxiv

Article Title: Programmable Lipid Nanoparticle Targeting via Corona Engineering

doi: 10.64898/2026.02.27.708523

Figure Lengend Snippet: (a) Fluorescence imaging and (b) flow cytometry analysis of LNP uptake in HEK293FT cells expressing cognate surface binders demonstrate that synthetic binder-dApoE fusion constructs enable receptor-specific transduction only in engineered cell lines, with minimal background in control cells. Scale bar, 100 µm. (c-d) scFv-based retargeting was evaluated using a CD19-specific scFv genetically fused to dApoE. While dApoE-CD19 scFv fusion LNPs mediated selective uptake in CD19-overexpressing HEK293FT cells relative to control cells, the magnitude of enhancement was modest and substantially lower than that achieved with wild-type ApoE.

Article Snippet: CD19 CAR mRNA was purchased from MedChemExpress and used to produce CAR-T cells generating LNPs.

Techniques: Fluorescence, Imaging, Flow Cytometry, Expressing, Construct, Transduction, Control

Journal: bioRxiv

Article Title: Programmable Lipid Nanoparticle Targeting via Corona Engineering

doi: 10.64898/2026.02.27.708523

Figure Lengend Snippet: In vivo generation of functional CAR T cells using detargeted and retargeted LNPs. (A) Schematic of CAR T cell generation strategy. (B) Representative microscope images showing CAR expression in T cells isolated from mice treated with CD5-conjugated or unconjugated LNPs (0.5 mg/kg RNA) with or without dApoE precoating (25× relative to LNP mRNA) or haPCSK9 pretreatment (40 µg per mouse, administered 15 min before LNP injection). Scale bar, 50 µm. (C) Flow cytometry quantification of CAR+ T cells following treatment with CD5-conjugated or unconjugated LNPs. (D) CAR expression in hepatocytes from treated animals, showing reduced liver CAR+ cells with detargeting strategies. (E) T-cell targeting ratio showing the ratio of T-cell transduction over liver transduction, normalized to LNP only (F) Functional killing assay showing cytotoxicity of engineered CAR T cells against CD19- and fLuc-overexpressing HEK293FT cells at 24h of co-culture. Cytotoxicity is calculated as the percent decrease in the luminescence signal compared to control. All data in this figure are mean ± SEM (n=3).

Article Snippet: CD19 CAR mRNA was purchased from MedChemExpress and used to produce CAR-T cells generating LNPs.

Techniques: In Vivo, Functional Assay, Microscopy, Expressing, Isolation, Injection, Flow Cytometry, Transduction, Co-Culture Assay, Control

Journal: Molecular Therapy Oncology

Article Title: Co-editing of NKG2A and FAS increases long-term cytotoxic capacity and persistence of CAR NK cells

doi: 10.1016/j.omton.2026.201126

Figure Lengend Snippet: NKG2A-edited CAR NK cells eliminate CD19-negative target cells resistant to conventional CAR NK cells (A) Specific lysis of 721.45 CD19 KO cells by engineered NK cells (left) or CAR NK cells (right). Cells were co-cultured for 4–5 h at an effector to target ratio of 5:1. (B) Schematic of repetitive stimulation assay. 0.1 × 10 6 NK or CAR NK cells were seeded at day 0, and 0.1 × 10 6 CD19-positive 721.45 cells (target cells) were added every 2–3 days for 14 days. IL-2 was added 2×/week until day 12. Target cells added on day 7 were labeled with PKH26 and on day 14 with PKH67. Data shown in (C–H) were generated using this setup. (C) Expression of CD19 (histograms) and relative MFI of CD19 on total target cells (live CD56 − ) and representative flow cytometry plots on day 14 after co-culture with NK (top) or CAR NK (bottom) cells. Relative MFI is calculated by dividing by MFI of CD19 on target cells cultured alone. (D) Representative expression of PKH26 (target cells added day 7) and PKH67 (target cells added day 14) on total target cells (live CD56 − ) after co-culture with NK or CAR NK cells for 14 days. (E and F) Frequency of remaining live PKH26 + target cells (added on day 7) on day 14 after co-culture with NK cells (E) or CAR NK cells (F). Target cells alone (721.45 only) were not included in the statistical analysis. (G and H) Frequency of remaining live PKH67 + target cells (added on day 14 and co-cultured for 4–5 h before readout) after co-culture with NK cells (G) or CAR NK cells (H). Target cells alone (721.45 only) were not included in the statistical analysis. (A–H) n = 6–9 (individual donors). One-way ANOVA. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: EBV-transformed LCLs from allogeneic human fetal liver tissue were generated by isolating CD19 + cells by magnetic-activated cell sorting with CD19 beads (Miltenyi, Cat. 130-050-301) and then infected with EBV B95-8, produced as previously described at a multiplicity of infection (MOI) of 0.05–0.15.

Techniques: Lysis, Cell Culture, Labeling, Generated, Expressing, Flow Cytometry, Co-Culture Assay

Journal: Molecular Therapy Oncology

Article Title: Co-editing of NKG2A and FAS increases long-term cytotoxic capacity and persistence of CAR NK cells

doi: 10.1016/j.omton.2026.201126

Figure Lengend Snippet: Increased cytotoxicity of NKG2A-edited CAR NK cells against CD19-negative BCP-ALL PDX (A) CD19 and (B) HLA-E expression of BCP-ALL PDX by flow cytometry. (C) Specific lysis of PDX1 and PDX2 after co-culture with control NK cells (NK mock) or CAR NK cells (CAR mock). Co-culture at E:T ratio 5:1 for 4 h. (D) Specific lysis of PDX1 after 4 h co-culture at 1:1 E:T ratio. (E) Data from (D) shown as relative increase in killing of engineered CAR NK cells compared with controls (CAR mock). (F–H) Specific lysis of PDX2 after 20 h co-culture at 5:1 E:T ratio with NK (F) or CAR NK (G) cells. (H) Data from (G) shown as relative increase in killing of engineered CAR NK cells compared with controls (CAR mock). (C) n = 3 (individual donors), paired t test. (D–G) n = 3 (individual donors), repeated measures one-way ANOVA. Engineered NK cells shown in this figure were generated from bulk NK cells (not NKG2A + NK cells) as the starting population. ∗ p < 0.05.

Article Snippet: EBV-transformed LCLs from allogeneic human fetal liver tissue were generated by isolating CD19 + cells by magnetic-activated cell sorting with CD19 beads (Miltenyi, Cat. 130-050-301) and then infected with EBV B95-8, produced as previously described at a multiplicity of infection (MOI) of 0.05–0.15.

Techniques: Expressing, Flow Cytometry, Lysis, Co-Culture Assay, Control, Generated

Journal: Nucleic Acids Research

Article Title: CFP1-mediated H3K4me3 broad domains control early B cell lineage fate determination

doi: 10.1093/nar/gkag125

Figure Lengend Snippet: CFP1-mediated broad H3K4me3 peaks promote lineage-specific gene expression in pro-B cells. ( A ) Volcano plot of DEGs (absolute fold change > 1.5 and Padj < 0.05) between Cfp1 -deficient and control pro-B cells. ( B ) Box plot comparing mRNA levels of significantly down-regulated and up-regulated genes. TPM, transcripts per million. ( C ) Average H3K4me3 signal (TSS ± 3 kb) of genes significantly down-regulated, with no difference (stable), or up-regulated upon Cfp1 loss. ( D ) Fraction of genes with significant H3K4me3 signal loss (down) or gain (up) at promoters among genes that are significantly down-regulated or up-regulated in expression in Cfp1 -deficient pro-B cells. ( E ) Violin plot comparing H3K4me3 peak widths in control cells between up-regulated and down-regulated genes upon Cfp1 loss. ( F ) Frequency of genes with different H3K4me3 peak widths among those with significantly altered expression upon Cfp1 deletion. ( G ) Volcano plot of DEGs in Cfp1 -deficient versus control pro-B cells, stratified by broad (left) and narrow (right) H3K4me3 peaks at promoters. ( H ) Bar plots showing enriched GO terms for genes that are down-regulated upon Cfp1 deletion and possess broad H3K4me3 domains at promoters. ( I ) H3K4me3 and RNA-seq profiles for Dntt, Il7r, Cd19, Ebf1, Blnk, Vpreb1 , and Igll1 genes in control and Cfp1 -deficient pro-B cells. Mann–Whitney U-tests were used for statistical analysis in (B) and (E). **** P < 0.0001.

Article Snippet: Cells were purified using CD19 MicroBeads (Miltenyi, 130-121-301) via magnetic cell sorting, followed by CD11b negative selection.

Techniques: Gene Expression, Control, Expressing, RNA Sequencing, MANN-WHITNEY