bluuebunny/crossref_metadata_2025 huggingface dataset:
Figure S2 from ERBB4-Mediated Signaling Is a Mediator of Resistance to PI3K and BTK Inhibitors in B-cell Lymphoid Neoplasms
Authors: Arribas Alberto J. , Napoli Sara , Cascione Luciano , Barnabei Laura , Sartori Giulio , Cannas Eleonora , Gaudio Eugenio , Tarantelli Chiara , Mensah Afua A. , Spriano Filippo , Zucchetto Antonella , Rossi Francesca M. , Rinaldi Andrea , Castro de Moura Manuel , Jovic Sandra , Bordone Pittau Roberta , Stathis Anastasios , Stussi Georg , Gattei Valter , Brown Jennifer R. , Esteller Manel , Zucca Emanuele , Rossi Davide , Bertoni Francesco
Published: March 2024

(A) Cell viability for the combination of idelalisib and the CD20 blocking antibody rituximab in parental and resistant by MTT assay (72h). Bars correspond to the mean of two independent experiments. Error bars represent standard deviation of the mean. Table contains p-values from a moderated t-test comparing each combination to idelalisib as single agent. The benefit of the combination was assessed both as synergism according to the Chou-Talalay combination index (CI, synergistic: CI<0.9, additive: CI~1, antagonistic: CI>1) (B) (2) and as potency (x-axis, synergistic: potency>0.5, additive: 01, additive: 0
Figure S17 from ERBB4-Mediated Signaling Is a Mediator of Resistance to PI3K and BTK Inhibitors in B-cell Lymphoid Neoplasms
Authors: Arribas Alberto J. , Napoli Sara , Cascione Luciano , Barnabei Laura , Sartori Giulio , Cannas Eleonora , Gaudio Eugenio , Tarantelli Chiara , Mensah Afua A. , Spriano Filippo , Zucchetto Antonella , Rossi Francesca M. , Rinaldi Andrea , Castro de Moura Manuel , Jovic Sandra , Bordone Pittau Roberta , Stathis Anastasios , Stussi Georg , Gattei Valter , Brown Jennifer R. , Esteller Manel , Zucca Emanuele , Rossi Davide , Bertoni Francesco
Published: March 2024

Sensitivity to ibrutinib was evaluated by MTT assay (72h). Karpas1718 parental (top) or resistant (bottom) were exposed to 100nM decitabine (A), 5µM tazemetostat (B), or DMSO (control, black) given concomitantly (continuous lines) or 72 hours before ibrutinib (pre DMSO in dashed black or pre decitabine/tazemetostat in dashed red). Error bars correspond to standard deviation of the mean. Data derived from two independent experiments. Statistically significant differences were evaluated by moderated t-test comparing each treatment to control (DMSO).
Figure S14 from ERBB4-Mediated Signaling Is a Mediator of Resistance to PI3K and BTK Inhibitors in B-cell Lymphoid Neoplasms
Authors: Arribas Alberto J. , Napoli Sara , Cascione Luciano , Barnabei Laura , Sartori Giulio , Cannas Eleonora , Gaudio Eugenio , Tarantelli Chiara , Mensah Afua A. , Spriano Filippo , Zucchetto Antonella , Rossi Francesca M. , Rinaldi Andrea , Castro de Moura Manuel , Jovic Sandra , Bordone Pittau Roberta , Stathis Anastasios , Stussi Georg , Gattei Valter , Brown Jennifer R. , Esteller Manel , Zucca Emanuele , Rossi Davide , Bertoni Francesco
Published: March 2024

Karpas1718 parental (A-B), SP53 (C-D), OCILY10 (C, E) and JEKO1-Cas9-ERBB4 (F-G) cells were exposed with DMSO (grey), idelalisib (red) or idelalisib + lapatinib (yellow) for 2-weeks. Expression levels of let-7c and miR-29c miRNAs (real-time PCR); and surface ERBB4 and intracellular HBEGF (both by FACS), were evaluated. RQ values calculated by the DDCt method. MFI for median fluorescence intensity. Expression levels of let-7c and miR-29c miRNAs (A), and surface ERBB4 and intracellular HBEGF (B) in Karpas1718 parental cells exposed with DMSO (grey), idelalisib (red) or idelalisib + lapatinib (yellow). Expression levels of let-7c and miR-29c miRNAs (C), and surface ERBB4 and intracellular HBEGF in SP53 (D) and OCILY10 (E) cells exposed with DMSO (grey), idelalisib (red) or idelalisib + lapatinib (yellow). Expression levels of surface ERBB4 and intracellular HBEGF in JEKO1-CNT (scramble sgRNA, F) and JEKO1-ERBB4 (sgRNA-ERBB4, G) cells exposed with DMSO (grey), idelalisib (red) or idelalisib + lapatinib (yellow). Data derived from two independent experiments; error bars represent standard deviation of the mean. * for p<0.05 from a t-test.
Figure S11 from ERBB4-Mediated Signaling Is a Mediator of Resistance to PI3K and BTK Inhibitors in B-cell Lymphoid Neoplasms
Authors: Arribas Alberto J. , Napoli Sara , Cascione Luciano , Barnabei Laura , Sartori Giulio , Cannas Eleonora , Gaudio Eugenio , Tarantelli Chiara , Mensah Afua A. , Spriano Filippo , Zucchetto Antonella , Rossi Francesca M. , Rinaldi Andrea , Castro de Moura Manuel , Jovic Sandra , Bordone Pittau Roberta , Stathis Anastasios , Stussi Georg , Gattei Valter , Brown Jennifer R. , Esteller Manel , Zucca Emanuele , Rossi Davide , Bertoni Francesco
Published: March 2024

Protein levels of ERBB4 was measured by FACS (A) and immunoblotting (B) in JEKO1 cells engineered to increase the expression of ERBB. Drug sensitivity for idelalisib (C) and ibrutinib (D) was evaluated in the different clones of engineered JEKO1. (A) Representative plot of two independent experiments. Boxplot and density plot shows the median MFI values of the two replicates: negative control (black), JEKO1-CNT (control, grey) and JEKO1-ERBB4 (sgRNA-erbb4, red). Results are presented from Welch Two Sample test performed on MFI values of the two experiments. (B) Immunoblotting for ERBB4 in JEKO1-CNT (control, grey) and JEKO1-ERBB4 (sgRNA-erbb4, red). Representative image of two independent experiments. Data on the barplot represents the mean of two experiments, errors bars represent standard deviation of the mean. * for p<0.05 from a t-test. (C-D) Cell viability was evaluated by MTT assay (72h). JEKO1-CNT (control, blue) and JEKO1-ERBB4 (ERBB4-sgRNA3 in red) cells were stimulated with PBS (left panel) or HBEGF (30ng/mL, right panel) for 4 hours and the exposed to increasing concentrations of idelalisib (C) or ibrutinib (D). Error bars correspond to standard deviation of the mean. Data derived from two independent experiments. Statistically significant differences were evaluated by moderated t-test comparing each treatment to control. * for p<0.05.
Figure S10 from ERBB4-Mediated Signaling Is a Mediator of Resistance to PI3K and BTK Inhibitors in B-cell Lymphoid Neoplasms
Authors: Arribas Alberto J. , Napoli Sara , Cascione Luciano , Barnabei Laura , Sartori Giulio , Cannas Eleonora , Gaudio Eugenio , Tarantelli Chiara , Mensah Afua A. , Spriano Filippo , Zucchetto Antonella , Rossi Francesca M. , Rinaldi Andrea , Castro de Moura Manuel , Jovic Sandra , Bordone Pittau Roberta , Stathis Anastasios , Stussi Georg , Gattei Valter , Brown Jennifer R. , Esteller Manel , Zucca Emanuele , Rossi Davide , Bertoni Francesco
Published: March 2024

Expression levels of crucial factors are correlated with resistance to idelalisib. HBEGF (A) and ERBB4 (B) expression levels are inversely correlated with idelalisib sensitivity. Conversely expression of let-7c (D) and miR-29c (F) microRNAs is associated with sensitivity to idelalisib. Expression and sensitivity data were analyzed from a previous publication of our group in a panel of 34 B cell lymphoma cell lines (4). Cell lines were split in two groups based on higher or lower values than the median expression of the corresponding gene or miRNA. Mean of idelalisib IC50 were calculated for these two groups and compared by t-test.
Figure S7 from Genetic Silencing of AKT Induces Melanoma Cell Death via mTOR Suppression
Authors: Parkman Gennie L. , Turapov Tursun , Kircher David A. , Burnett William J. , Stehn Christopher M. , O'Toole Kayla , Culver Katie M. , Chadwick Ashley T. , Elmer Riley C. , Flaherty Ryan , Stanley Karly A. , Foth Mona , Lum David H. , Judson-Torres Robert L. , Friend John E. , VanBrocklin Matthew W. , McMahon Martin , Holmen Sheri L.
Published: March 2024

Supplementary Figure S7 shows chemical structures of all pharmacological agents utilized in study, including MK2206, GSK2141795, GDC-0941, BYL719, RapaLink-1, GSK650394, GDC-0068, GDC-0084, and INY-03-041.
Figure S6 from Genetic Silencing of AKT Induces Melanoma Cell Death via mTOR Suppression
Authors: Parkman Gennie L. , Turapov Tursun , Kircher David A. , Burnett William J. , Stehn Christopher M. , O'Toole Kayla , Culver Katie M. , Chadwick Ashley T. , Elmer Riley C. , Flaherty Ryan , Stanley Karly A. , Foth Mona , Lum David H. , Judson-Torres Robert L. , Friend John E. , VanBrocklin Matthew W. , McMahon Martin , Holmen Sheri L.
Published: March 2024

Supplementary Figure S6 shows that INY-03-041 leads to a substantial effect on melanoma cell proliferation but it is not as potent as siAKT1-3. A, Cell confluency assay of MTG001 and MTG004 cell lines treated with DMSO control, GDC0068 (1µM), INY-03-041 (1µM), GDC0941 (1µM), Lenalidomide (1µM), or siAKT1-3 (50nM). Treatment with INY-03-041 led to decreased cell proliferation (P < 0.0001 in each cell line), but did not lead to cell death, similar to siAKT1-3 in MTG001 and MTG004. Lenalidomide, a recruiter of the E3 ubiquitin ligase Cereblon, conjugated to GDC0068 to form INY-03-041, was used as a negative control for the off-target effects of INY-03-041. Error bars indicate standard error of the mean of triplicate wells. B, Immunoblotting of cell lysates treated with the DMSO control, MK2206 (2.5µM), GDC0068 (1µM), INY-03-041 (500nM), GSK0394 (3µM), GDC0941 (1µM), BYL719 (1µM), siAKT1-3 (50nM) at 22 h, and siAKT1-3 (50nM) for 24 h revealed complete knockdown of P-AKT (Ser473) and total AKT by siAKT1-3 at 24 h, as well as knockdown of P-PRAS40, P-S6 kinase, and P-4EBP1. INY-03-041 displayed a 77% reduction in total AKT compared to the 89% reduction by siAKT1-3. C, Time-course immunoblotting of cell lysates treated with varying concentrations of INY-03-041 demonstrated an incomplete total protein knockdown of individual AKT paralogs. D, Densitometry quantification of time-course immunoblotting found in C.
Figure S5 from Genetic Silencing of AKT Induces Melanoma Cell Death via mTOR Suppression
Authors: Parkman Gennie L. , Turapov Tursun , Kircher David A. , Burnett William J. , Stehn Christopher M. , O'Toole Kayla , Culver Katie M. , Chadwick Ashley T. , Elmer Riley C. , Flaherty Ryan , Stanley Karly A. , Foth Mona , Lum David H. , Judson-Torres Robert L. , Friend John E. , VanBrocklin Matthew W. , McMahon Martin , Holmen Sheri L.
Published: March 2024

Supplemental Figure 5 shows that caspase 3 and 8 inhibition protects melanoma cells from siAKT1-3-mediated apoptosis. A, Cell confluency assay upon treatment with siAKT1-3 versus co-treatment with caspase inhibitors. Co-treatment of A375 and MTG004 cells with Z-VAD-FMK, Z-DEVD-FMK, and Z-IETD-FMK inhibitors protected cells against siAKT1-3-mediated cell death. B, Incuycte® Cytotox Red assay comparing siCtrl, siAKT1-3, siCtrl + each caspase inhibitor, and siAKT1-3 + caspase inhibitors. C, Incuycte® Caspase 3/7 assay comparing siCtrl, siAKT1-3, siCtrl + each caspase inhibitor, and siAKT1-3 + caspase inhibitors. The percentage of dead cells stained with Cytotox red and caspase 3/7 reagents increased only in the siAKT1-3 group, indicating complete rescue with caspase 3, 8, or pan inhibition.
Figure S2 from Genetic Silencing of AKT Induces Melanoma Cell Death via mTOR Suppression
Authors: Parkman Gennie L. , Turapov Tursun , Kircher David A. , Burnett William J. , Stehn Christopher M. , O'Toole Kayla , Culver Katie M. , Chadwick Ashley T. , Elmer Riley C. , Flaherty Ryan , Stanley Karly A. , Foth Mona , Lum David H. , Judson-Torres Robert L. , Friend John E. , VanBrocklin Matthew W. , McMahon Martin , Holmen Sheri L.
Published: March 2024

Supplementary Figure S2 shows the classification of MTG001 and MTG004. Immunoblotting of MTG001 and MTG004 PDX-derived cell lines showed similar levels of P-AKT (Ser473), total AKT, P-ERK (Thr202/Tyr204), ERK1/2, P-Ribosomal S6 Kinase (Ser235/236), and S6 Kinase. GAPDH was used as a loading control. The MTG001 cells exhibited higher levels of total PTEN. Analysis of RNA sequencing data demonstrated that both MTG001 and MTG004 express wildtype PTEN.
Supplementary Table S1 from Palazestrant (OP-1250), A Complete Estrogen Receptor Antagonist, Inhibits Wild-type and Mutant ER-positive Breast Cancer Models as Monotherapy and in Combination
Authors: Parisian Alison D. , Barratt Susanna A. , Hodges-Gallagher Leslie , Ortega Fabian E. , Peña Guadalupe , Sapugay Judevin , Robello Brandon , Sun Richard , Kulp David , Palanisamy Gopinath S. , Myles David C. , Kushner Peter J. , Harmon Cyrus L.
Published: March 2024

PROSeq analysis of gene expression changes following treatment of MCF7 and CAMA-1 cells with ER-alpha ligands.
Supplementary Figure S6 from Palazestrant (OP-1250), A Complete Estrogen Receptor Antagonist, Inhibits Wild-type and Mutant ER-positive Breast Cancer Models as Monotherapy and in Combination
Authors: Parisian Alison D. , Barratt Susanna A. , Hodges-Gallagher Leslie , Ortega Fabian E. , Peña Guadalupe , Sapugay Judevin , Robello Brandon , Sun Richard , Kulp David , Palanisamy Gopinath S. , Myles David C. , Kushner Peter J. , Harmon Cyrus L.
Published: March 2024

Combination of OP-1250 with palbociclib or ribociclib enhances suppression of cell cycle-related gene expression. A, t-SNE plot of MCF7 xenograft samples from vehicle, low-dose OP-1250, and palbociclib monotherapy and combination treatment groups. B, C, Heatmap showing expression of genes associated with the E2 late gene signature and E2F gene signature of n = 3 MCF7 xenograft tumors from OP-1250 and palbociclib low dose monotherapy and combination therapy groups. Red indicates high expression relative to vehicle and blue indicates low expression. D, Comparison of log2-fold change values for 14 cell cycle-related genes between combination therapy (pink) and monotherapies (blue and purple for 1 mg/kg OP-1250 and 25 mg/kg palbociclib, respectively). Stacked bars represent the combined effect of the monotherapies according to the Bliss independence model. E, F, Heatmaps showing expression of genes associated with the G2/M and E2 late gene signatures of n = 3 MCF7 xenograft tumors from OP-1250 and ribociclib low dose monotherapy and combination therapy groups. Red indicates high expression relative to vehicle and blue indicates low expression. G, Venn diagram showing overlap in dysregulated genes relative to vehicle of 1 mg/kg OP-1250 + 25 mg/kg palbociclib and 1 mg/kg OP-1250 + 25 mg/kg ribociclib MCF7 xenograft tumors. H, Heatmap of genes associated with the kinetochore metaphase signaling pathway, the most enriched pathway (p-value 6.15×10-7) in 676 genes changed in both OP-1250 + CDK4/6 inhibitor treatment groups. Genes are color-coded by gene expression relative to vehicle, with red indicating high expression and blue indicating low expression. OP-1250 1 mg/kg treatment groups from two xenograft studies were averaged.
Supplementary Figure S5 from Palazestrant (OP-1250), A Complete Estrogen Receptor Antagonist, Inhibits Wild-type and Mutant ER-positive Breast Cancer Models as Monotherapy and in Combination
Authors: Parisian Alison D. , Barratt Susanna A. , Hodges-Gallagher Leslie , Ortega Fabian E. , Peña Guadalupe , Sapugay Judevin , Robello Brandon , Sun Richard , Kulp David , Palanisamy Gopinath S. , Myles David C. , Kushner Peter J. , Harmon Cyrus L.
Published: March 2024

Addition of OP-1250 to CDK4/6 inhibitors enhances tumor suppression and prolongs animal survival in ER+ breast cancer xenograft models. A, B, Percent change in tumor volume of individual animals in listed treatment groups at day 29 of the MCF7 xenograft study, normalized to day 1 tumor volume. C, D, Percent change in tumor volume of individual animals in listed treatment groups at day 29 of the ST941 xenograft study, normalized to day 1 tumor volume. E, F, Kaplan–Meier graphs of animal survival over time in listed treatment groups of mice implanted with the ESR1Y537S ST941 PDX model. Dosing ended on day 29 of the study, and animals were sacrificed when tumor volume reached 2000 mm3. G, H, Mean and SEM tumor volume over the full study interval with monotherapy and combination treatments in the ESR1Y537S ST941 PDX breast cancer model, with the dotted line representing tumor stasis. Dosing ended on day 29, as indicated by the dashed line.
Table S6 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

Differentially expressed proteins identified from the total proteomic analysis in SUDHL5 relative to VAL DLBCL cells (untreated).
Table S4 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

Raw data for kinase binding affinity assay with NBI-961.
Table S12 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

KEA3 analysis that used the Cheng kinase-substrate interaction network in SUDHL5 and RIVA treated with NBI-961 relative to DMSO vehicle control using the top 25 proteins from each with a signficant loss of phosphorylation (FDR < 0.05; log2 fold change less than or equal to -1).
Table S10 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

Differentially expressed proteins identified from the phospho-proteomic analysis in SUDHL5 treated with NBI-961 relative to DMSO vehicle control.
Figure S8 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

Doxorubicin alone as a positive control induces apoptosis. Stacked bar graph of cell lines untreated, treated with DMSO or doxorubicin (DOX) shows percent viable cells and cells in early or late apoptosis. Data are shown as mean ± SEM from three independent experiments. Adjusted P-values as determined by a two-way ANOVA analysis with Tukey’s multiple test correction for comparisons to DMSO treated control. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001
Figure S6 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

Original blot images from Figure 5H in the main text. Uncropped blot images of Figure 5H. Samples were run on duplicate gels and each duplicate blot was immunoblotted with either a mixture of anti-NEK2 and anti-ACTIN primary antibodies (left panels) or a mixture of anti-p-AKT, anti-AKT, and anti-ACTIN primary antibodies (right panels). The blots were imaged in both the Dylight 650 and 800 channels and shown as the respective image (NEK2 or p-AKT from 650 in top panels, and AKT and ACTIN from 800 in lower panels). All blots were resolved with a mixture of secondary antibodies Dylight 650 and Dylight 800 and imaged using the BioRad ChemiDoc MP as described in the methods.
Supplementary Figure S1 from Dual CAR-T Cells Targeting CD19 and CD37 Are Effective in Target Antigen Loss B-cell Tumor Models
Authors: Imai Kanae , Takeuchi Yuki , Terakura Seitaro , Okuno Shingo , Adachi Yoshitaka , Osaki Masahide , Umemura Koji , Hanajiri Ryo , Shimada Kazuyuki , Murata Makoto , Kiyoi Hitoshi
Published: March 2024

Supplemental Figure S1. CD19 and CD37 expression of EBV-LCL. EBV-LCL was stained with CD19-PE and CD37-FITC mAb, respectively. Grey tinted area indicated isotype control staining.
Figure S4 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

Original blot images from Figure 4 in the main text. Uncropped blot images of (A) Figure 4A; (B) Figure 4C; (C) Figure 4G; and (D) Figure 4H. Each blot was imaged in both the Dylight 650 and 800 channels and shown as the respective image (NEK2 from 650 in top panels and ACTIN from 800 in lower panels). All blots were immunoblotted with a mixture of anti-NEK2 and anti-ACTIN primary antibodies, resolved with a mixture of secondary antibodies Dylight 650 and Dylight 800 and imaged using the BioRad ChemiDoc MP as described in the methods.
Figure S11 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

NBI-961 causes chemosensitization in a subset of DLBCL cell lines. Bar graphs showing percent cell viability of benign B cells (GM16113) and DLBCL cell lines after 96 h NBI-961 treatment alone or in combination with doxorubicin (DOX) or vincristine (VCR). Adjusted P-values as determined by a one-way ANOVA with Dunnett’s multiple test correction for comparisons to either DOX or VCR only treated controls.
Figure S1 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

Synthesis of NBI-961. Reagents and Conditions: a) NaOMe, MeOH 0 °C – rt. b) PPh3, DIAD, DCM 0 °C - rt. c) LDA, dry THF, -78 °C – rt. d) Na2CO3, Pd(dppf)Cl2.DCM, 4:1 THF:H2O. e) 7M NH3 in MeOH, NH4OH, 78 °C. f) K2CO3, Pd(dppf)Cl2.DCM, DMF, 80 °C.
Supplementary Legends from Raludotatug Deruxtecan, a CDH6-Targeting Antibody–Drug Conjugate with a DNA Topoisomerase I Inhibitor DXd, Is Efficacious in Human Ovarian and Kidney Cancer Models
Authors: Suzuki Hirokazu , Nagase Shotaro , Saito Chiemi , Takatsuka Atsuko , Nagata Motoko , Honda Kokichi , Kaneda Yuki , Nishiya Yumi , Honda Tomoyo , Ishizaka Tomomichi , Nakamura Kensuke , Nakada Takashi , Abe Yuki , Agatsuma Toshinori
Published: March 2024

Supplementary Figure and Table Legends
Figure S2 from Raludotatug Deruxtecan, a CDH6-Targeting Antibody–Drug Conjugate with a DNA Topoisomerase I Inhibitor DXd, Is Efficacious in Human Ovarian and Kidney Cancer Models
Authors: Suzuki Hirokazu , Nagase Shotaro , Saito Chiemi , Takatsuka Atsuko , Nagata Motoko , Honda Kokichi , Kaneda Yuki , Nishiya Yumi , Honda Tomoyo , Ishizaka Tomomichi , Nakamura Kensuke , Nakada Takashi , Abe Yuki , Agatsuma Toshinori
Published: March 2024

CDH6 expression in cell lines.
Figure S9 from Improving Intracellular Delivery of an Antibody–Drug Conjugate Targeting Carcinoembryonic Antigen Increases Efficacy at Clinically Relevant Doses In Vivo
Authors: Nessler Ian , Rubahamya Baron , Kopp Anna , Hofsess Scott , Cardillo Thomas M. , Sathyanarayan Nalini , Donnell Jennifer , Govindan Serengulam V. , Thurber Greg M.
Published: March 2024

Quantifying CEACAM6 expression on LS174T cells. Human Fc binding beads (Bangs Laboratories) were incubated with AF488 labelled Tinurilimab biosimilar. Correlation of median fluorescence intensity with number of binding sites per bead was generated and the slope used to calculate the number of CEACAM6 receptors per LS174T cell. Approximately 24,857 antibodies/cell were bound, so CEACAM6 expression was estimated at 49,714 receptors per LS174T cell assuming bivalent binding.
Figure S6 from Improving Intracellular Delivery of an Antibody–Drug Conjugate Targeting Carcinoembryonic Antigen Increases Efficacy at Clinically Relevant Doses In Vivo
Authors: Nessler Ian , Rubahamya Baron , Kopp Anna , Hofsess Scott , Cardillo Thomas M. , Sathyanarayan Nalini , Donnell Jennifer , Govindan Serengulam V. , Thurber Greg M.
Published: March 2024

In vitro toxicity curves. LS174T cells were seeded in 96 well plates and then incubated with a serial dilution of either LG or XAb-SN-38 for 6 days with media replacement every 24 hours. The toxicity curves are presented in either protein (A) or payload (B) incubation concentration. Although XAb-SN-38 provided more efficacy in vivo, the two ADCs resulted in similar in vitro potency.
Figure S5 from Improving Intracellular Delivery of an Antibody–Drug Conjugate Targeting Carcinoembryonic Antigen Increases Efficacy at Clinically Relevant Doses In Vivo
Authors: Nessler Ian , Rubahamya Baron , Kopp Anna , Hofsess Scott , Cardillo Thomas M. , Sathyanarayan Nalini , Donnell Jennifer , Govindan Serengulam V. , Thurber Greg M.
Published: March 2024

CEA Antibody Internalization Rate. LS174T cells were incubated continuously with 20 nM of antibody, and the internalization rate was measured as previously reported1. For both antibodies, total and surface fluorescence increased over the course of the experiment. The approximately tenfold difference in MFI between XAb and hMN-14 binding was due to the different flow cytometer voltage settings on the days the experiments were run(A). The increase in surface fluorescence is likely due to LS174T cell packing that can increase the time for binding equilibrium and delay surface fluorescence plateau. The main difference between XAb and hMN-14 conjugates was the differing internalization rates, consistent with literature reports2. The internalized fluorescence for XAb increased throughout the 8-hour incubation while hMN-14 internalized fluorescence tapered off towards the end of the continuous incubation. The data were used to determine the time integral of the surface fluorescence and plotted against the internal fluorescence (B). A linear fit was used to calculate the slope which corresponds to ke.
Figure S4 from Improving Intracellular Delivery of an Antibody–Drug Conjugate Targeting Carcinoembryonic Antigen Increases Efficacy at Clinically Relevant Doses In Vivo
Authors: Nessler Ian , Rubahamya Baron , Kopp Anna , Hofsess Scott , Cardillo Thomas M. , Sathyanarayan Nalini , Donnell Jennifer , Govindan Serengulam V. , Thurber Greg M.
Published: March 2024

HiSep XAb-SN-38. HPLC detection at 360 nm, SN-38 absorbance region, with 98.7% associated with the peak corresponding to the antibody. Peak at 8.123 mm is due to unremoved SN-38.
Fig. S20 from Radiation Synergizes with IL2/IL15 Stimulation to Enhance Innate Immune Activation and Antitumor Immunity
Authors: Li Xuefeng , Huntoon Kristin , Wang Yifan , Lee DaeYong , Dong Shiyan , Antony Abin , Walkey Carl , Kim Betty Y.S. , Jiang Wen
Published: March 2024

Fig. S20
Figure S3 from Improving Intracellular Delivery of an Antibody–Drug Conjugate Targeting Carcinoembryonic Antigen Increases Efficacy at Clinically Relevant Doses In Vivo
Authors: Nessler Ian , Rubahamya Baron , Kopp Anna , Hofsess Scott , Cardillo Thomas M. , Sathyanarayan Nalini , Donnell Jennifer , Govindan Serengulam V. , Thurber Greg M.
Published: March 2024

SE-HPLC Plot XAb-SN-38. HPLC detection at 360 nm, SN-38 absorbance region, with 100% associated with the peak corresponding to the antibody.
Fig. S14 from Radiation Synergizes with IL2/IL15 Stimulation to Enhance Innate Immune Activation and Antitumor Immunity
Authors: Li Xuefeng , Huntoon Kristin , Wang Yifan , Lee DaeYong , Dong Shiyan , Antony Abin , Walkey Carl , Kim Betty Y.S. , Jiang Wen
Published: March 2024

Fig. S14
Fig. S1 from Radiation Synergizes with IL2/IL15 Stimulation to Enhance Innate Immune Activation and Antitumor Immunity
Authors: Li Xuefeng , Huntoon Kristin , Wang Yifan , Lee DaeYong , Dong Shiyan , Antony Abin , Walkey Carl , Kim Betty Y.S. , Jiang Wen
Published: March 2024

Fig. S1
Data from Genetic Silencing of AKT Induces Melanoma Cell Death via mTOR Suppression
Authors: Parkman Gennie L. , Turapov Tursun , Kircher David A. , Burnett William J. , Stehn Christopher M. , O'Toole Kayla , Culver Katie M. , Chadwick Ashley T. , Elmer Riley C. , Flaherty Ryan , Stanley Karly A. , Foth Mona , Lum David H. , Judson-Torres Robert L. , Friend John E. , VanBrocklin Matthew W. , McMahon Martin , Holmen Sheri L.
Published: March 2024

Abstract
Aberrant activation of the PI3K–AKT pathway is common in many cancers, including melanoma, and AKT1, 2 and 3 (AKT1–3) are bona fide oncoprotein kinases with well-validated downstream effectors. However, efforts to pharmacologically inhibit AKT have proven to be largely ineffective. In this study, we observed paradoxical effects following either pharmacologic or genetic inhibition of AKT1–3 in melanoma cells. Although pharmacological inhibition was without effect, genetic silencing of all three AKT paralogs significantly induced melanoma cell death through effects on mTOR. This phenotype was rescued by exogenous AKT1 expression in a kinase-dependent manner. Pharmacological inhibition of PI3K and mTOR with a novel dual inhibitor effectively suppressed melanoma cell proliferation in vitro and inhibited tumor growth in vivo. Furthermore, this single-agent–targeted therapy was well-tolerated in vivo and was effective against MAPK inhibitor–resistant patient-derived melanoma xenografts. These results suggest that inhibition of PI3K and mTOR with this novel dual inhibitor may represent a promising therapeutic strategy in this disease in both the first-line and MAPK inhibitor–resistant setting.
Data from Palazestrant (OP-1250), A Complete Estrogen Receptor Antagonist, Inhibits Wild-type and Mutant ER-positive Breast Cancer Models as Monotherapy and in Combination
Authors: Parisian Alison D. , Barratt Susanna A. , Hodges-Gallagher Leslie , Ortega Fabian E. , Peña Guadalupe , Sapugay Judevin , Robello Brandon , Sun Richard , Kulp David , Palanisamy Gopinath S. , Myles David C. , Kushner Peter J. , Harmon Cyrus L.
Published: March 2024

Abstract
The estrogen receptor (ER) is a well-established target for the treatment of breast cancer, with the majority of patients presenting as ER-positive (ER+). Endocrine therapy is a mainstay of breast cancer treatment but the development of resistance mutations in response to aromatase inhibitors, poor pharmacokinetic properties of fulvestrant, agonist activity of tamoxifen, and limited benefit for elacestrant leave unmet needs for patients with or without resistance mutations in ESR1, the gene that encodes the ER protein. Here we describe palazestrant (OP-1250), a novel, orally bioavailable complete ER antagonist and selective ER degrader. OP-1250, like fulvestrant, has no agonist activity on the ER and completely blocks estrogen-induced transcriptional activity. In addition, OP-1250 demonstrates favorable biochemical binding affinity, ER degradation, and antiproliferative activity in ER+ breast cancer models that is comparable or superior to other agents of interest. OP-1250 has superior pharmacokinetic properties relative to fulvestrant, including oral bioavailability and brain penetrance, as well as superior performance in wild-type and ESR1-mutant breast cancer xenograft studies. OP-1250 combines well with cyclin-dependent kinase 4 and 6 inhibitors in xenograft studies of ER+ breast cancer models and effectively shrinks intracranially implanted tumors, resulting in prolonged animal survival. With demonstrated preclinical efficacy exceeding fulvestrant in wild-type models, elacestrant in ESR1-mutant models, and tamoxifen in intracranial xenografts, OP-1250 has the potential to benefit patients with ER+ breast cancer.
Data from Raludotatug Deruxtecan, a CDH6-Targeting Antibody–Drug Conjugate with a DNA Topoisomerase I Inhibitor DXd, Is Efficacious in Human Ovarian and Kidney Cancer Models
Authors: Suzuki Hirokazu , Nagase Shotaro , Saito Chiemi , Takatsuka Atsuko , Nagata Motoko , Honda Kokichi , Kaneda Yuki , Nishiya Yumi , Honda Tomoyo , Ishizaka Tomomichi , Nakamura Kensuke , Nakada Takashi , Abe Yuki , Agatsuma Toshinori
Published: March 2024

Abstract
Cadherin-6 (CDH6) is expressed in several cancer types, but no CDH6-targeted therapy is currently clinically available. Here, we generated raludotatug deruxtecan (R-DXd; DS-6000), a novel CDH6-targeting antibody–drug conjugate with a potent DNA topoisomerase I inhibitor, and evaluated its properties, pharmacologic activities, and safety profile. In vitro pharmacologic activities and the mechanisms of action of R-DXd were assessed in serous-type ovarian cancer and renal cell carcinoma cell lines. In vivo pharmacologic activities were evaluated with several human cancer cell lines and patient-derived xenograft mouse models. The safety profile in cynomolgus monkeys was also assessed. R-DXd exhibited CDH6 expression-dependent cell growth-inhibitory activity and induced tumor regression in xenograft models. In this process, R-DXd specifically bound to CDH6, was internalized into cancer cells, and then translocated to the lysosome. The DXd released from R-DXd induced the phosphorylation of Chk1, a DNA damage marker, and cleaved caspase-3, an apoptosis marker, in cancer cells. It was also confirmed that the DXd payload had a bystander effect, passing through the cell membrane and impacting surrounding cells. The safety profile of R-DXd was favorable and the highest non-severely toxic dose was 30 mg/kg in cynomolgus monkeys. R-DXd demonstrated potent antitumor activity against CDH6-expressing tumors in mice and an acceptable safety profile in monkeys. These findings indicate the potential of R-DXd as a new treatment option for patients with CDH6-expressing serous-type ovarian cancer and renal cell carcinoma in a clinical setting.
Data from Improving Intracellular Delivery of an Antibody–Drug Conjugate Targeting Carcinoembryonic Antigen Increases Efficacy at Clinically Relevant Doses In Vivo
Authors: Nessler Ian , Rubahamya Baron , Kopp Anna , Hofsess Scott , Cardillo Thomas M. , Sathyanarayan Nalini , Donnell Jennifer , Govindan Serengulam V. , Thurber Greg M.
Published: March 2024

Abstract
Solid tumor antibody–drug conjugates (ADC) have experienced more clinical success in the last 5 years than the previous 18-year span since the first ADC approval in 2000. While recent advances in protein engineering, linker design, and payload variations have played a role in this success, high expression and readily internalized targets have also been crucial to solid tumor therapy. However, these factors are also paradoxically connected to poor tissue penetration and lower efficacy. Previous work shows that potent ADCs can benefit from slower internalization under subsaturating doses to improve tissue penetration and increase tumor response. In contrast, faster internalization is predicted to increase efficacy under higher, tumor saturating doses. In this work, the intracellular delivery of SN-38 conjugated to an anti-carcinoembryonic antigen (anti-CEA) antibody (Ab) is increased by coadministering a noncompeting (cross-linking) anti-CEA Ab to improve efficacy in a colorectal carcinoma animal model. The SN-38 payload enables broad tumor saturation with clinically-tolerable doses, and under these saturating conditions, using a second CEA receptor cross-linking Ab yields faster internalization, which increases tumor killing efficacy. Our spheroid results show indirect bystander killing can also occur, but the more efficient direct cell killing from targeted intracellular payload release drives a greater tumor response. These results provide a strategy to increase therapeutic effectiveness with improved intracellular delivery under tumor saturating doses with the potential to expand the ADC target repertoire.
Abstract A015: Predicting efficacious drugs and drug-biomarker relationships for targeted glioblastoma treatment
Authors: Maeser Danielle , Gruener Robert , Galvin Robert , Huang Stephanie , Koga Tomo , Grigore Florina
Published: March 2024

Glioblastoma multiforme (GBM) is the deadliest, most heterogeneous, and common brain cancer in adults. Despite major advancements in neurosurgery as well as chemotherapy and radiotherapy techniques, overall prognosis has changed little over the decades. Not only is there an urgent need to identify efficacious therapeutics but there is also a great need to pair these therapeutics with biomarkers that can help tailor treatment to the right patient populations given the heterogeneous nature of the disease. Here, we implement several machine learning and statistical based pipelines for drug and biomarker discovery, leveraging previous work building patient drug response models using high-throughput cell line drug screening data as well as Bayesian network structure learning. Through these discovery pipelines, we identified multiple agents of interest for GBM, including MEK inhibitors (MEKis). MEKis were consistently predicted to be effective against GBM in multiple cross platform (microarray and RNA-sequencing) patient and patient derived xenograft (PDX) datasets. Additionally, they were experimentally shown to be more efficacious in tumor samples than standard of care agents (Temozolomide and Carmustine). We also predicted PHGDH gene expression levels to be causally associated with MEKi efficacy, which increased tumor sensitivity to a MEKi (Trametinib) during experimental testing in GBM cell lines with knockdown. These findings support our drug and biomarker discovery pipelines as an effective strategy to unite multiple cross platform and cross species datasets to help achieve GBM precision medicine. They also demonstrate that patient PHGDH levels can help inform Trametinib response in the fight against GBM.

Citation Format: Danielle Maeser, Robert Gruener, Robert Galvin, Stephanie Huang, Tomo Koga, Florina Grigore. Predicting efficacious drugs and drug-biomarker relationships for targeted glioblastoma treatment [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr A015.
Abstract A033: Murine CNS and bone marrow distribution of the aurora A kinase inhibitor alisertib: Pharmacokinetics and exposure at the sites of efficacy and toxicity
Authors: Oh Juhee , Power Erica A. , Zhang Wenjuan , Daniels David J. , Elmquist William F.
Published: March 2024

Important challenges in developing drugs that target central nervous system (CNS) tumors include overcoming barriers for CNS delivery and reducing systemic side effects. Alisertib, an aurora A kinase inhibitor, has been examined for treatment of several CNS tumors in preclinical and clinical studies. In this study, we investigated the distribution of alisertib into the CNS, the site of efficacy for brain tumors, and into the bone marrow, the site of dose-limiting toxicity leading to myelosuppression. Mechanisms influencing site-specific distribution, such as active transport mediated by the efflux proteins, p-glycoprotein (P-gp) and breast cancer resistance protein (Bcrp), were examined. Alisertib exposure to the brain in wild-type mice was less than 1% of that in the plasma and was evenly distributed throughout various brain regions and the spinal cord. Studies using transporter knockout mice and pharmacological inhibition show that alisertib CNS distribution is influenced by P-gp, but not Bcrp. Conversely, upon systemic administration, alisertib distribution to the bone marrow occurred rapidly, was not significantly limited by efflux transporters, and reached higher concentrations than in the CNS. This study demonstrates that, given an equivalent distributional driving force exposure in plasma, the exposure of alisertib in the brain is significantly less than that in the bone marrow, suggesting that targeted delivery may be necessary to guarantee therapeutic efficacy with minimal risk for adverse events. Therefore, these data suggest that, to improve the therapeutic index when using alisertib for brain tumors, a localized regional delivery, such as convection-enhanced delivery, may be warranted.

Citation Format: Juhee Oh, Erica A. Power, Wenjuan Zhang, David J. Daniels, William F. Elmquist. Murine CNS and bone marrow distribution of the aurora A kinase inhibitor alisertib: Pharmacokinetics and exposure at the sites of efficacy and toxicity [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr A033.
Abstract B022: Interplay between the cellular dntp pool and dna double-strand break repair: a potential therapeutic target for refractory glioblastoma
Authors: Monroe Dominique , Usoro Edidiong , Williams Arilyn , Aycox Shelby , Steinbeck Emily , Daddacha Waaqo
Published: March 2024

Glioblastoma (GBM) is a deadly brain tumor resistant to double-strand break (DSB)-inducing therapeutics due to dysregulated DNA-repair. Ionizing radiation (IR) and some chemotherapeutics strategically target cancer cells by causing DNA-DSBs, resulting in cell death if left insufficiently repaired. Homologous Recombination (HR) is an error-free DSB-repair process consisting of proteins that are often dysregulated in GBM, causing overactive DNA-repair, therapy resistance, and poor patient outcomes. Proteins involved in the HR-repair process have been extensively investigated; however, therapeutic resistance remains common. Here, we provide an alternative approach to target DSB-resistant GBM by modulating the dNTP pool instead of only targeting DNA-repair proteins. Our goal is to understand the interplay between dNTP pools and DSB-repair. dNTPs are a necessary constituent to repair DSBs; however, the role of the dNTP pool in DNA damage repair is not fully understood. Previous research shows that the dNTP pool remains unchanged after inducing DSBs, highlighting the importance of a tightly regulated dNTP pool. Our results show that increasing the dNTP pool significantly increased apoptosis and reduced cancer cell viability after IR. We found that a higher cellular dNTP pool impaired HR-mediated DSB-repair by delaying DNA end-resection in HR. Filling this gap in our knowledge may provide new avenues to sensitize therapy-resistant cancers, enhancing the efficacy of DSB-inducing therapies.

Citation Format: Dominique Monroe, Edidiong Usoro, Arilyn Williams, Shelby Aycox, Emily Steinbeck, Waaqo Daddacha. Interplay between the cellular dntp pool and dna double-strand break repair: a potential therapeutic target for refractory glioblastoma [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr B022.
Abstract B032: Designing arrays for the mouse head to facilitate in vivo studies of Tumor Treating Fields (TTFields) treatment of glioblastoma
Authors: Zbidat Sewar , Blatt Roni , Sellevoll Mariell , Gabay Martin , Schlachet Inbar , Cahal Shay , Davidi Shiri , Tzchori Itai , El-Mabhouh Amal , Haber Adi , Giladi Moshe , Palti Yoram
Published: March 2024

Background: Tumor Treating Fields (TTFields) therapy is a noninvasive, anticancer treatment delivered by two pairs of arrays placed on the skin surrounding the tumor location. Based on efficacy and safety data from phase 3 global clinical studies, TTFields therapy is FDA-approved and CE marked for treatment of glioblastoma (GBM), the most common malignant brain tumor. In vivo studies of TTFields in GBM models, which are mostly based on mice, are complicated by the relatively small size of the mouse head and corresponding limited surface area to apply the arrays. This challenge has thus far limited in vivo research on TTFields treatment in GBM. Materials and Methods: To identify a method for efficient delivery of TTFields to the mouse brain, we examined the ability of different array layouts to accommodate the geometries of the mouse head while minimally impacting head movement, taking into account that it may not be feasible to place two pairs of arrays on the head. The properties of different adhesive tapes, specifically their ability to attach to the skin, was also investigated. Finally, the ability of array layouts to deliver therapeutic intensities to the desired region was validated using simulations and direct electric field measurements. Results: Assessments showed an optimal layout consisted of arrays such that one of the pair was on the mouse head, and the other on the mouse torso; and the arrays for placement on the head divided into two smaller disks, A thin, transparent adhesive tape was identified that allowed the arrays to be securely attached to the skin, but was easily removed without leaving residues. Validation studies using simulations demonstrated that the array layout met the minimum treatment requirements (field intensity ≥1 V/cm RMS; current ≥50 mA), and facilitated TTFields usage for ≥75% of each day. Conclusions: These data show that TTFields treatment can be effectively delivered to the heads of mice by utilizing carefully designed arrays. Our newly developed mouse head arrays are a flexible construct, that adheres strongly to the skin, and enables efficient electric field delivery to the mouse brain. These improvements in the delivery of TTFields treatment to mice will facilitate research of TTFields as a therapy for GBM.

Citation Format: Sewar Zbidat, Roni Blatt, Mariell Sellevoll, Martin Gabay, Inbar Schlachet, Shay Cahal, Shiri Davidi, Itai Tzchori, Amal El-Mabhouh, Adi Haber, Moshe Giladi, Yoram Palti. Designing arrays for the mouse head to facilitate in vivo studies of Tumor Treating Fields (TTFields) treatment of glioblastoma [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr B032.