Journal: Translational Cancer Research

Article Title: In vitro characterization of a novel anti-GD2/ROR1 bispecific antibody exhibiting tumor cell killing efficacy in neuroblastoma cross-cancer activity in ROR1-positive triple-negative breast cancer

doi: 10.21037/tcr-2026-0723

Figure Lengend Snippet: Molecular design schematic and purification gel image of bispecific antibody G/R-001. (A) Molecular design diagram of the light chain and heavy chain of bispecific antibody G/R-001. (B) Schematic diagram of GD2/ROR1 bispecific antibody G/R-001. (C) SDS-PAGE gel image of the bispecific antibody under reducing or non-reducing conditions. The left shows the SDS-PAGE gel image under reducing conditions, with two bands in the lane-the upper band representing the heavy chain and the lower band representing the light chain; the right shows the SDS-PAGE gel image under non-reducing conditions, with one band in the lane representing the intact antibody. CMV, cytomegalovirus; G/R-001, anti-GD2/ROR1 bispecific antibody; GD2, disialoganglioside 2; H, heavy chain; IgG, immunoglobulin G; L, light chain; ROR1, receptor tyrosine kinase-like orphan receptor 1; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; VH, variable domain of heavy chain; VK, variable domain of light chain.

Article Snippet: Isotype IgG (Cat#: HY-P99001, MedChemExpress, Princeton, NJ, USA), naxitamab (anti-GD2) (Cat#: HY-P99206, MedChemExpress), zilovertamab (anti-ROR1) (Cat#: HY-P99201, MedChemExpress), and GD2 × ROR1 BsAb were transiently expressed in HEK293 cells using the OPM-293 system (Cat#: 91070-010, OPM, Shanghai, China).

Techniques: Purification, SDS Page, Polyacrylamide Gel Electrophoresis

Journal: Translational Cancer Research

Article Title: In vitro characterization of a novel anti-GD2/ROR1 bispecific antibody exhibiting tumor cell killing efficacy in neuroblastoma cross-cancer activity in ROR1-positive triple-negative breast cancer

doi: 10.21037/tcr-2026-0723

Figure Lengend Snippet: ELISA detection of the binding activity of bispecific antibody G/R-001 to GD2 and ROR1. GD2 or ROR1 antigens were coated, and G/R-001, zilovertamab, naxitamab, and isotype IgG antibodies were added to the ELISA plate at concentrations of 0–150 nM with 2-fold serial dilutions. After secondary antibody incubation, absorbance values were detected at 450 nM wavelength by a microplate reader, curves were plotted, and EC 50 values were calculated. (A) Binding affinity of G/R-001 and naxitamab to GD2, with EC 50 values of 12.1 and 11.9 nM, respectively; (B) Binding affinity of G/R-001 and zilovertamab to ROR1, with EC 50 values of 3.5 and 3.1 nM, respectively. EC 50 , half maximal effective concentration; ELISA, enzyme-linked immunosorbent assay; G/R-001, anti-GD2/ROR1 bispecific antibody; GD2, disialoganglioside 2; IgG, immunoglobulin G; OD, optical density; ROR1, receptor tyrosine kinase-like orphan receptor 1.

Article Snippet: Isotype IgG (Cat#: HY-P99001, MedChemExpress, Princeton, NJ, USA), naxitamab (anti-GD2) (Cat#: HY-P99206, MedChemExpress), zilovertamab (anti-ROR1) (Cat#: HY-P99201, MedChemExpress), and GD2 × ROR1 BsAb were transiently expressed in HEK293 cells using the OPM-293 system (Cat#: 91070-010, OPM, Shanghai, China).

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Activity Assay, Incubation, Concentration Assay

Journal: Translational Cancer Research

Article Title: In vitro characterization of a novel anti-GD2/ROR1 bispecific antibody exhibiting tumor cell killing efficacy in neuroblastoma cross-cancer activity in ROR1-positive triple-negative breast cancer

doi: 10.21037/tcr-2026-0723

Figure Lengend Snippet: Flow cytometry detection of the binding activity of bispecific antibody G/R-001 to GD2/ROR1 positive cells. GD2 − /ROR1 − HEK293 cells were used as a negative control. G/R-001, zilovertamab, naxitamab, and isotype IgG at 10 nM concentration showed significant binding activity to GD2 − /ROR1 + MDA-MB-468 cells, GD2 + /ROR1 − LAN-1 cells, and GD2 + /ROR1 + MHH-NB-11 cells. FITC, fluorescein isothiocyanate; G/R-001, anti-GD2/ROR1 bispecific antibody; GD2, disialoganglioside 2; IgG, immunoglobulin G; ROR1, receptor tyrosine kinase-like orphan receptor 1.

Article Snippet: Isotype IgG (Cat#: HY-P99001, MedChemExpress, Princeton, NJ, USA), naxitamab (anti-GD2) (Cat#: HY-P99206, MedChemExpress), zilovertamab (anti-ROR1) (Cat#: HY-P99201, MedChemExpress), and GD2 × ROR1 BsAb were transiently expressed in HEK293 cells using the OPM-293 system (Cat#: 91070-010, OPM, Shanghai, China).

Techniques: Flow Cytometry, Binding Assay, Activity Assay, Negative Control, Concentration Assay

Journal: Translational Cancer Research

Article Title: In vitro characterization of a novel anti-GD2/ROR1 bispecific antibody exhibiting tumor cell killing efficacy in neuroblastoma cross-cancer activity in ROR1-positive triple-negative breast cancer

doi: 10.21037/tcr-2026-0723

Figure Lengend Snippet: LDH assay detection of the in vitro killing activity of bispecific antibody G/R-001 against GD2/ROR1 positive neuroblastoma and other cells. Isotype IgG, naxitamab, zilovertamab, naxitamab + zilovertamab, and G/R-001 antibodies (50 µL) at different concentrations (0–100.0 nM) were added to the experimental wells and cultured for 48 h. Following the LDH kit procedure, absorbance was read at 490 nm using a microplate reader. (A) HEK293 cells (GD2 − /ROR1 − ), (B) LAN-1 cells (GD2 + /ROR1 − ), (C) MDA-MB-468 cells (GD2 − /ROR1 + ), and (D) MHH-NB-11 cells (GD2 + /ROR1 + ). With increasing concentrations, Naxitamab, Zilovertamab, Naxitamab+Zilovertamab, and G/R-001 antibodies showed significantly increased toxicity against target-positive cells, with G/R-001 demonstrating the most significant cytotoxicity. G/R-001, anti-GD2/ROR1 bispecific antibody; GD2, disialoganglioside 2; IgG, immunoglobulin G; LDH, lactate dehydrogenase; ROR1, receptor tyrosine kinase-like orphan receptor 1.

Article Snippet: Isotype IgG (Cat#: HY-P99001, MedChemExpress, Princeton, NJ, USA), naxitamab (anti-GD2) (Cat#: HY-P99206, MedChemExpress), zilovertamab (anti-ROR1) (Cat#: HY-P99201, MedChemExpress), and GD2 × ROR1 BsAb were transiently expressed in HEK293 cells using the OPM-293 system (Cat#: 91070-010, OPM, Shanghai, China).

Techniques: Lactate Dehydrogenase Assay, In Vitro, Activity Assay, Cell Culture

Journal: Chemmedchem

Article Title: Novel Para ‐Phenylenediamine‐Based Derivatives as Receptor Tyrosine Kinase‐like Orphan Receptor 1 (ROR1) Inhibitors: An In Vitro Preliminary Characterization

doi: 10.1002/cmdc.202500247

Figure Lengend Snippet: Known chemical structures of ROR1 inhibitors.

Article Snippet: Recombinant human ROR1 protein (Fc Tag) and recombinant human ROR1 protein (aa 453‐783, His&GST Tag) were purchased from Elabscience (Texas, USA).

Techniques:

Journal: Chemmedchem

Article Title: Novel Para ‐Phenylenediamine‐Based Derivatives as Receptor Tyrosine Kinase‐like Orphan Receptor 1 (ROR1) Inhibitors: An In Vitro Preliminary Characterization

doi: 10.1002/cmdc.202500247

Figure Lengend Snippet: Structure‐based drug design approach leading to the para‐ phenylenediamine derivatives acting as ROR1 inhibitors.

Article Snippet: Recombinant human ROR1 protein (Fc Tag) and recombinant human ROR1 protein (aa 453‐783, His&GST Tag) were purchased from Elabscience (Texas, USA).

Techniques:

Journal: Chemmedchem

Article Title: Novel Para ‐Phenylenediamine‐Based Derivatives as Receptor Tyrosine Kinase‐like Orphan Receptor 1 (ROR1) Inhibitors: An In Vitro Preliminary Characterization

doi: 10.1002/cmdc.202500247

Figure Lengend Snippet: CETSA identifies ROR1 protein in the binding with 17 in JeKo‐1 cells. A) CETSA western blots and B) corresponding quantitative analysis of band intensity. 17 group was treated with 17 (40 μM) for 2 h and then subjected to 5 min incubation at the indicated temperature (45–58 °C). Ctrl group was treated with vehicle (0.1% DMSO) only in the same conditions. ROR1 level at 25 °C was set at 100%. Densitometry‐based quantification of western blotting signals was calculated by first normalizing to α ‐tubulin levels in individual samples. Statistical analyses were performed comparing the compound 17 group to the Ctrl group across different temperature points. Results are shown as mean ± SD from three independent experiments. ** and *** denote respectively p < 0.01 and p < 0.001 versus Ctrl.

Article Snippet: Recombinant human ROR1 protein (Fc Tag) and recombinant human ROR1 protein (aa 453‐783, His&GST Tag) were purchased from Elabscience (Texas, USA).

Techniques: Binding Assay, Western Blot, Incubation

Journal: Chemmedchem

Article Title: Novel Para ‐Phenylenediamine‐Based Derivatives as Receptor Tyrosine Kinase‐like Orphan Receptor 1 (ROR1) Inhibitors: An In Vitro Preliminary Characterization

doi: 10.1002/cmdc.202500247

Figure Lengend Snippet: Molecular effects of 17 on ROR1 pathways in JeKo‐1 cells. For each western blotting analysis, three independent experiments were performed, and the SD is expressed by error bars. The relative fold change versus untreated cells, set as 1, is shown in the graph. *, **, and *** denote respectively p < 0.05, p < 0.01, and p < 0.001 versus Ctrl.

Article Snippet: Recombinant human ROR1 protein (Fc Tag) and recombinant human ROR1 protein (aa 453‐783, His&GST Tag) were purchased from Elabscience (Texas, USA).

Techniques: Western Blot

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: (A) Map of retroviral constructs used to generate ROR1 CAR and control T cells. TM = transmembrane. hIgG4 = human immunoglobulin G4. scFv = single chain variable fragment. (B) Percent change in body weight in BALB/c mice (left) and representative flow cytometric plots showing frequency of CD8+CD45.1+ donor T cells of live cells and frequency of CD8+tCD19+ cells of CD45.1+ donor T cells 7 days post-transfer (right). (C) Percent change in body weight in BALB/c mice. n=4 mice per group. Two-way ANOVA with Tukey’s post-test (ROR1 CAR-T vs control T at Day 7, 9, 11: p<0.00001). (D) Representative flow cytometric plots showing expression of tCD19 transduction marker and PD-1 on CD8+CD45.1+ donor T cells 4 days post-transfer (left) and summary of absolute number and PD-1 median fluorescence intensity (MFI) on CD45.1+CD8+tCD19+ control or ROR1 CAR-T cells 4 days post-transfer (right). n=4 mice per group. Unpaired Student’s two-way t-test (Abs #: spleen, p=0.0004; BM, p=0.0002; PD-1: spleen, p=0.0007; BM, p=0.0007; blood, p=0.000008). (E) White blood cell (WBC), lymphocyte, red blood cell (RBC), and platelet (PLT) counts in peripheral blood. n=4 mice per group. Two-way ANOVA with Tukey’s post-test (ROR1 CAR-T vs control T: Lymphocytes Day 14, p<0.005; RBC Day 10,14, p<0.00001; PLT Day 14, p<0.005). (F) Representative H&E stains of spleen and femur (BM) 14 days post-transfer (left) and histopathology scoring of control or ROR1 CAR-T cell-treated tissues (right). n=4 mice per group. Two-way ANOVA with Sidak post-test (Spleen Day 8,10,14, p<0.00001; BM Day 10, p=0.004; BM Day 14, p<0.00001). Data are representative of 5 independent experiments. All data are presented as the mean values ± SEM. See also Table S1 and Figures S1-S2.

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Retroviral, Construct, Control, Expressing, Transduction, Marker, Fluorescence, Histopathology

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: (A) Percent change in body weight in BALB/c mice lymphodepleted (LD) as indicated. n=4 mice per group. Two-way ANOVA with Tukey’s post-test (ROR1 CAR-T vs control T: 500 R Day 7, 9, p<0.00001; 200 mg/kg cyclophosphamide (Cy) Day 7, p=0.0004; 200 mg/kg Cy Day 9, p<0.00001). (B) Frequency of CD8+CD45.1+tCD19+ ROR1 CAR-T cells in peripheral blood of BALB/c mice receiving 100 R, 500 R, 100 mg/kg Cy, or 200 mg/kg Cy for LD. n=4 mice per group. (C-E) Percent change in body weight (C), frequency (D) and PD-1 and TIM3 expression (E) on CD8+CD45.1+tCD19+ donor T cells in peripheral blood of BALB/c mice irradiated 500 R and treated as indicated. n=4 mice per group. Two-way ANOVA with Tukey’s post-test (1 M CAR-T vs control T: Day 7, p=0.0003; Day 9,18, p<0.00001; 0.5 M CAR-T vs control T: Day 20, p<0.00001; Day 24, p=0.0005; Day 27, p=0.0044). Data are representative of 2 independent experiments. All data are presented as the mean values ± SEM. See also Figure S3.

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Control, Expressing, Irradiation

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: (A) Percent change in body weight (left), RBC count in peripheral blood (middle), and representative pictures of spleens 9 days post-transfer (right) from B6 Ror1fl/fl (WT) or EIIα-Cre+Ror1fl/fl (KO) mice treated with 500 R and control or ROR1 CAR-T cells. n=3 mice per group. Left: two-way ANOVA with Tukey post-test (WT+CAR-T vs. KO+CAR-T: Day 7,8,9, p<0.00001). Middle: one-way ANOVA with Tukey post-test (p=0.0003). (B) Percent change in body weight (left), RBC count in peripheral blood (middle), and representative pictures of spleens 15 days post-transfer (right) from WT>WT or ROR1-KO>WT BM chimeric mice treated with 500 R and control or ROR1 CAR-T cells. n=3 mice per group. Left: two-way ANOVA with Tukey post-test (WT>WT CAR-T vs. KO>WT CAR-T: n.s. = not significant). Middle: one-way ANOVA with Tukey post-test. (C) Percent change in body weight (left), RBC count in peripheral blood (middle), and representative pictures of spleens 40 days post-transfer (right) from WT>WT or WT>ROR1-KO BM chimeric mice treated with 500 R and control or ROR1 CAR-T cells. n=3 mice per group. Left: two-way ANOVA with Tukey post-test (WT>WT CAR-T vs. WT>KO CAR-T: Day 7, p=0.00013; Day 9, p=0.0045; Day 10,11, p<0.00001). Middle: one-way ANOVA with Tukey post-test (p=0.0044). Data are representative of 2 independent experiments. All data are presented as the mean values ± SEM.

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Control

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: (A) qPCR analysis of Ror1 expression in sorted BM progenitors (left) and BM stromal cells (right) from BALB/c mice left untreated or irradiated 500 R 48 hr prior to euthanasia. Ror1 expression was normalized to Actb expression and expressed relative to Ror1 expression in E10.5 embryos. OBL = osteoblasts. MSC = mesenchymal stem cells. EC = endothelial cells. n=3 mice per group. (B) ELISA analysis of IFNγ production by CD8+ control or ROR1 CAR-T cells co-cultured for 48 hr with primary MSC derived from WT or ROR1-KO femurs and expanded in vitro for 14 days. Data are summarized from 3 independent experiments. Unpaired two-way Student’s t-test (p=0.0041). (C) Absolute number of cells in BM flushed from femurs and tibia of control or ROR1 CAR-T cell-treated mice 9 days post-transfer (left) and number of MSC, pre-B cell, granulocyte/macrophage, erythrocyte, and megakaryocyte colonies formed after culture of BM cells from femurs of control or ROR1 CAR-T cell-treated BALB/c mice collected 9 days post-transfer (right). n=3 mice per group. Unpaired two-way Student’s t-test (# BM cells, p=0.0004; MSC, p=0.0004; Pre-B, p=0.0023). (D) ELISA analysis of IFNγ production by CD8+ control or ROR1 CAR-T cells co-cultured for 48 hr with CD45+ or CD45−splenic cells. Data are summarized from 3 independent experiments. Unpaired two-way Student’s t-test (p=0.0049). (E) qPCR analysis of Ror1 expression in splenic subpopulations sorted from BALB/c mice left untreated or irradiated 500 R 48 hr prior to euthanasia. n.d. = not detected. Data are summarized from 3 independent experiments. All data are presented as the mean values ± SEM.

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Expressing, Irradiation, Enzyme-linked Immunosorbent Assay, Control, Cell Culture, Derivative Assay, In Vitro

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: (A) Representative flow cytometric analysis of EpCAM expression on 4T1 tumor cells, MSC, osteoblasts (OBL), CD45−Ter119−PDGFRβ+ spleen cells, and CD45−Ter119−VE-cadherin+ spleen cells from BALB/c mice. Percent lysis of the indicated 51Cr-labeled tumor cells after 6 hr (top) or 24 hr (bottom) of co-culture with CD8+ untransduced (black), ROR1 CAR-T cells (gray), or EpCAM-inducible ROR1 CAR-T cells (open circles). (C) Intracellular cytokine analysis of tCD19 CAR marker and IFNγ expression in the indicated T cells co-cultured with tumor cells in the presence of Brefeldin A for the last 6 hr of culture. Data are representative of two independent experiments. See also Figures S4 and S5.

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Expressing, Lysis, Labeling, Co-Culture Assay, Marker, Cell Culture

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: (A) Percent change in body weight in BALB/c mice inoculated with 4T1-mROR1 tumors and treated as indicated. n=6–12 mice per group. Two-way ANOVA with Tukey post-test (ROR1-CAR vs. EpCAM-inducible ROR1 CAR: Day 4, p<0.0001; Day 7, p<0.0001; Day 10, p=0.0002). (B) RBC and PLT counts from peripheral blood of tumor-bearing mice treated as indicated. n=6–12 mice per group. Two-way ANOVA with Tukey post-test (ROR1-CAR vs. EpCAM-inducible ROR1 CAR: RBC Day 4,8,15,22:, p<0.0001; PLT Day 8 p=0.0114; PLT Day 15, 22, p<0.0001). (C) Representative pictures of spleens 10 days post-transfer from tumor-bearing BALB/c mice treated as indicated. (D) Tumor volume in BALB/c mice treated as indicated. n=6–12 mice per group. Two-way ANOVA with Tukey post-test (ROR1-CAR vs. UT: Day 14, p<0.0001; Day 17, p<0.0001; EpCAM-inducible ROR1 CAR vs. UT: Day 14, p<0.0001; Day 17, p<0.0001; ROR1-CAR vs EpCAM-inducible ROR1 CAR: n.s). (E) Survival of mice treated as indicated. n=6–12. Log-rank Mantel-Cox test (UT vs. ROR1 CAR, p=0.0003; UT vs. EpCAM-inducible ROR1 CAR, p=0.0016; ROR1 CAR vs. EpCAM-inducible ROR1 CAR, p=0.0001). (F) Representative flow cytometric analysis of ROR1 CAR and PD-1 expression on CD8+CD45.1+ donor T cells from mice treated with Cy and the indicated T cells 10 days post-transfer. (G) Summary of frequency of ROR1 CAR+, CAR+Ki67+, and CAR+PD-1+ T cells of CD8+CD45.1+ donor T cells from mice treated with Cy and the indicated T cells 10 days post-transfer. n=6–12 mice per group. Unpaired two-way Student’s t-test (Frequency: spleen, p<0.0001; BM, p<0.0001; PD-1: spleen, p=0.0004; BM, p=0.0003; Ki67: spleen, p<0.0001; BM, p<0.0001). Data are summarized from 2 independent experiments. All data are presented as the mean values ± SEM.

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Expressing

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: (A) Map of lentiviral constructs encoding CD19-Gal4VP64 synNotch Receptor, UAS inducible R11 ROR1 CAR, and constitutive R11 ROR1 CAR. (B) Raji tumor bioluminescence over time in NSG mice irradiated 250 R and treated with the indicated primary human T cells. n=6–7 per group. Two-way ANOVA with Tukey post-test (UT vs. ROR1 CAR-T: Day 13, 20, p<0.0001; UT vs. CD19-inducible ROR1 CAR-T: Day 13, 20, p<0.0001). (C) Summary of tEGFR CAR marker and PD-1 expression on the indicated donor CD8+CD45+ T cells in peripheral blood. n=4–7 per group. Two-way ANOVA with Tukey post-test (CAR: UT vs. ROR1 CAR, Day 3, 7, p<0.0001; ROR1 CAR vs. CD19-inducible ROR1 CAR, Day 3, p<0.0001; Day 7, n.s. PD-1: UT vs. ROR1 CAR, Day 3, 7, p<0.0001; ROR1 CAR vs. CD19-inducible ROR1 CAR, Day 3, p<0.0001; Day 7, n.s.). (D) Survival of tumor-bearing mice irradiated 250 R and treated with the indicated T cell groups. n=6–7 per group. Log-rank Mantel-Cox test (UT vs. ROR1 CAR, p=0.0007; UT vs. CD19-inducible ROR1 CAR, p=0.0007; ROR1 CAR vs. CD19-inducible ROR1 CAR, n.s.). Data are representative of 2 independent experiments. All data are presented as the mean values ± SEM. See also Figure S6.

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Construct, Irradiation, Marker, Expressing

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: (A) Percent change in body weight in NSG mice implanted with MDA-MB-231 tumors, irradiated 250 R and treated with the indicated primary human T cells. n=4 mice per group. Two-way ANOVA with Tukey post-test (ROR1-CAR vs. EpCAM-inducible ROR1 CAR: Day 13, 14, 19, p<0.0001). (B) RBC and PLT counts from peripheral blood of tumor-bearing mice treated as indicated 20 days post-T cell transfer. n=4 mice per group. Two-way ANOVA with Tukey post-test (RBC: ROR1 CAR vs. EpCAM-inducible ROR1 CAR, p=0.0426; UT vs. ROR1 CAR, p=0.0418. PLT: ROR1 CAR vs. EpCAM-inducible ROR1 CAR, p=0.0117; UT vs. ROR1 CAR, p=0.0442). (C and D) Tumor bioluminescence (C) and survival (D) of NSG mice treated as indicated. n=4 mice per group. Two-way ANOVA with Tukey post-test (ROR1 CAR vs. UT: Day 13, p=0.0167; Day 20, p<0.0001. EpCAM-inducible ROR1 CAR vs. UT: Day 13, p=0.0029; Day 20, p<0.0001. ROR1 CAR vs EpCAM-inducible ROR1 CAR: n.s). (E) Representative flow cytometric analysis of ROR1 CAR and Ki67 expression on CD8+CD45+ donor T cells from NSG mice irradiated 250 R and given indicated T cells 22 days post-transfer. (F) Summary of frequency of ROR1 CAR+, Ki67+, and PD-1+ T cells of CD8+CD45+ donor T cells from mice irradiated 250 R and given indicated T cells 22 days post-transfer. n=4 mice per group. Unpaired two-way Student’s t-test (Frequency: spleen, p<0.00001; BM, p<0.00001; tumor, n.s. Ki67: spleen, p<0.0001; BM, p<0.0001; tumor, n.s. PD-1: spleen, p<0.0001; BM, p<0.0001; tumor, n.s.). Data are representative of 2 independent experiments. All data are presented as the mean values ± SEM. See also Figures S7 and S8.

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Irradiation, Expressing

Journal: Cancer cell

Article Title: Logic-gated ROR1 chimeric antigen receptor expression rescues T cell-mediated toxicity to normal tissues and enables selective tumor targeting

doi: 10.1016/j.ccell.2019.02.003

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-human ROR1 APC , Miltenyi Biotec , Cat# 130–117-942, RRID:AB_2733449.

Techniques: Control, Histopathology, Recombinant, Cell Isolation, cDNA Synthesis, Plasmid Preparation, Software, SYBR Green Assay