bluuebunny/crossref_metadata_2025 huggingface dataset:
Supplementary Table 1 from Enhanced GAB2 Expression Is Associated with Improved Survival in High-Grade Serous Ovarian Cancer and Sensitivity to PI3K Inhibition
Authors: Davis Sally J. , Sheppard Karen E. , Anglesio Michael S. , George Joshy , Traficante Nadia , Fereday Sian , Intermaggio Maria P. , Menon Usha , Gentry-Maharaj Aleksandra , Lubinski Jan , Gronwald Jacek , Pearce Celeste Leigh , Pike Malcolm C. , Wu Anna , Kommoss Stefan , Pfisterer Jacobus , du Bois Andreas , Hilpert Felix , Ramus Susan J. , Bowtell David D.L. , Huntsman David G. , Pearson Richard B. , Simpson Kaylene J. , Campbell Ian G. , Gorringe Kylie L.
Published: April 2023

Nanostring data including clinical information
The sensitiviy of gefitinib in various NSCLC cell lines from Targeting Adenine Nucleotide Translocase-2 (ANT2) to Overcome Resistance to Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor in Non–Small Cell Lung Cancer
Authors: Jang Ji-Young , Kim Yong-goo , Nam Soo Jeong , Keam Bhumsuk , Kim Tae Min , Jeon Yoon Kyung , Kim Chul Woo
Published: April 2023

PC9, H1975, HCC827, and HCC827/GR cells were treated with gefitinib or DMSO as a control and subsequently incubated for an additional 24 h. Then, cell viability was measured using CCK8 assays and presented as a relative value compared to that of DMSO-treated cells. The values presented in all of the histograms are the mean values +/- SD.
Supplementary Figure Legends, Tables 1 - 6 from The Selective PI3K Inhibitor XL147 (SAR245408) Inhibits Tumor Growth and Survival and Potentiates the Activity of Chemotherapeutic Agents in Preclinical Tumor Models
Authors: Foster Paul , Yamaguchi Kyoko , Hsu Pin P. , Qian Fawn , Du Xiangnan , Wu Jianming , Won Kwang-Ai , Yu Peiwen , Jaeger Christopher T. , Zhang Wentao , Marlowe Charles K. , Keast Paul , Abulafia Wendy , Chen Jason , Young Jenny , Plonowski Artur , Yakes F. Michael , Chu Felix , Engell Kelly , Bentzien Frauke , Lam Sanh T. , Dale Stephanie , Yturralde Olivia , Matthews David J. , Lamb Peter , Laird A. Douglas
Published: April 2023

Supplemental Table S1: Kinase profile for XL147. Supplemental Table S2: Effects of XL147 on PI3K Pathway Signaling in MCF7 and PC-3 Cells. Supplemental Table S3: XL147 potency on cell proliferation by genotype. Supplemental Table S4: Effects of XL147 on Cell Cycle of MCF7 Cells. Supplemental Table S5: XL147 plasma concentrations, pharmacodynamics assays. Supplemental Table S6: Efficacy of XL147 in U-87 MG, A549, A2058, and WM-266-4 xenograft tumors. Supplemental Figure Legends (Figures S1 to S6).
Supplementary Figure Legends from The Selective Anaplastic Lymphoma Receptor Tyrosine Kinase Inhibitor ASP3026 Induces Tumor Regression and Prolongs Survival in Non–Small Cell Lung Cancer Model Mice
Authors: Mori Masamichi , Ueno Yoko , Konagai Satoshi , Fushiki Hiroshi , Shimada Itsuro , Kondoh Yutaka , Saito Rika , Mori Kenichi , Shindou Nobuaki , Soga Takatoshi , Sakagami Hideki , Furutani Takashi , Doihara Hitoshi , Kudoh Masafumi , Kuromitsu Sadao
Published: April 2023

PDF file - 68K
Supplementary Methods, Figure Legends 1-4 from Evasion Mechanisms to Igf1r Inhibition in Rhabdomyosarcoma
Authors: Abraham Jinu , Prajapati Suresh I. , Nishijo Koichi , Schaffer Beverly S. , Taniguchi Eri , Kilcoyne Aoife , McCleish Amanda T. , Nelon Laura D. , Giles Francis G. , Efstratiadis Argiris , LeGallo Robin D. , Nowak Brent M. , Rubin Brian P. , Malempati Suman , Keller Charles
Published: April 2023

Supplementary Methods, Figure Legends 1-4 from Evasion Mechanisms to Igf1r Inhibition in Rhabdomyosarcoma
Supplemental Figure 1 from The MDM2 Inhibitor AMG 232 Demonstrates Robust Antitumor Efficacy and Potentiates the Activity of p53-Inducing Cytotoxic Agents
Authors: Canon Jude , Osgood Tao , Olson Steven H. , Saiki Anne Y. , Robertson Rebecca , Yu Dongyin , Eksterowicz John , Ye Qiuping , Jin Lixia , Chen Ada , Zhou Jing , Cordover David , Kaufman Stephen , Kendall Richard , Oliner Jonathan D. , Coxon Angela , Radinsky Robert
Published: April 2023

Supplemental Figure 1. AMG 232 chemical structure, and 3-D model of AMG 232 bound to MDM2 protein.
Supplementary Figure S1 from TAS-116, a Highly Selective Inhibitor of Heat Shock Protein 90α and β, Demonstrates Potent Antitumor Activity and Minimal Ocular Toxicity in Preclinical Models
Authors: Ohkubo Shuichi , Kodama Yasuo , Muraoka Hiromi , Hitotsumachi Hiroko , Yoshimura Chihoko , Kitade Makoto , Hashimoto Akihiro , Ito Kenjiro , Gomori Akira , Takahashi Koichi , Shibata Yoshihiro , Kanoh Akira , Yonekura Kazuhiko
Published: April 2023

Supplementary Figure S1: Biodistribution of ganetespib, 17-DMAG, or SNX-5422 in athymic nude rats with established NCI-H1975 xenografts.
Fig. S1 from A miR-29b Byproduct Sequence Exhibits Potent Tumor-Suppressive Activities via Inhibition of NF-κB Signaling in KRAS-Mutant Colon Cancer Cells
Authors: Inoue Akira , Mizushima Tsunekazu , Wu Xin , Okuzaki Daisuke , Kambara Nanami , Ishikawa Sho , Wang Jiaqi , Qian Yamin , Hirose Haruka , Yokoyama Yuhki , Ikeshima Ryo , Hiraki Masayuki , Miyoshi Norikatsu , Takahashi Hidekazu , Haraguchi Naotsugu , Hata Taishi , Matsuda Chu , Doki Yuichiro , Mori Masaki , Yamamoto Hirofumi
Published: April 2023

Supplementary Figure S1. Effects of mimic-miR-29b-1-5p on cell proliferation.
Figure S4 from HIF2α-Targeted RNAi Therapeutic Inhibits Clear Cell Renal Cell Carcinoma
Authors: Wong So C. , Cheng Weijun , Hamilton Holly , Nicholas Anthony L. , Wakefield Darren H. , Almeida Aaron , Blokhin Andrei V. , Carlson Jeffrey , Neal Zane C. , Subbotin Vladimir , Zhang Guofeng , Hegge Julia , Bertin Stephanie , Trubetskoy Vladimir S. , Rozema David B. , Lewis David L. , Kanner Steven B.
Published: April 2023

In vivo trigger screen
Figure S1 from Acquired JHDM1D–BRAF Fusion Confers Resistance to FGFR Inhibition in FGFR2-Amplified Gastric Cancer
Authors: Sase Hitoshi , Nakanishi Yoshito , Aida Satoshi , Horiguchi-Takei Kana , Akiyama Nukinori , Fujii Toshihiko , Sakata Kiyoaki , Mio Toshiyuki , Aoki Masahiro , Ishii Nobuya
Published: April 2023

Cell growth inhibition by 5-FU and oxaliplatin in SNU-16 parental cell and four AZD-R clones.
Supplementary Materials from Predicting Novel Therapies and Targets: Regulation of Notch3 by the Bromodomain Protein BRD4
Authors: Villar-Prados Alejandro , Wu Sherry Y. , Court Karem A. , Ma Shaolin , LaFargue Christopher , Chowdhury Mamur A. , Engelhardt Margaret I. , Ivan Cristina , Ram Prahlad T. , Wang Ying , Baggerly Keith , Rodriguez-Aguayo Cristian , Lopez-Berestein Gabriel , Yang Shyh-Ming , Maloney David J. , Yoshioka Makoto , Strovel Jeffrey W. , Roszik Jason , Sood Anil K.
Published: April 2023

File contains synthesis of CN210, sequences for siRNAs, supplementary tables 1 to 3, supplementary figures 1 through 8 and figure legends.
Supplementary Figure 2 from Emodin Inhibits Breast Cancer Growth by Blocking the Tumor-Promoting Feedforward Loop between Cancer Cells and Macrophages
Authors: Iwanowycz Stephen , Wang Junfeng , Hodge Johnie , Wang Yuzhen , Yu Fang , Fan Daping
Published: April 2023

Emodin inhibits STAT6 and CEBP/β signaling in TAMs
Supplementary Methods from RG7386, a Novel Tetravalent FAP-DR5 Antibody, Effectively Triggers FAP-Dependent, Avidity-Driven DR5 Hyperclustering and Tumor Cell Apoptosis
Authors: Brünker Peter , Wartha Katharina , Friess Thomas , Grau-Richards Sandra , Waldhauer Inja , Koller Claudia Ferrara , Weiser Barbara , Majety Meher , Runza Valeria , Niu Huifeng , Packman Kathryn , Feng Ningping , Daouti Sherif , Hosse Ralf J. , Mössner Ekkehard , Weber Thomas G. , Herting Frank , Scheuer Werner , Sade Hadassah , Shao Cuiying , Liu Bin , Wang Peng , Xu Gary , Vega-Harring Suzana , Klein Christian , Bosslet Klaus , Umaña Pablo
Published: April 2023

Supplementary information on cell maintenance, and antibody selection and characterization
Supplementary Figures 1-3 with legends from Dual mTORC1/2 Inhibition as a Novel Strategy for the Resensitization and Treatment of Platinum-Resistant Ovarian Cancer
Authors: Musa Fernanda , Alard Amandine , David-West Gizelka , Curtin John P. , Blank Stephanie V. , Schneider Robert J.
Published: April 2023

Supplementary Figure S1: Growth inhibitory 50% (GI50) determination of single agent everolimus, PP242 and carboplatin using OVCAR-3 cells; Supplementary Figure S2: mTORC1 and mTORC1/2 inhibition sensitizes platinum resistant SKOV3 cells to carboplatin-mediated growth inhibition; Supplementary Figure S3: qRT-PCR analysis of select mRNAs in mTORC1 and mTORC1/2 inhibited cells with and without carboplatin treatment.
Supplementary Figure S1, Table S1, and Table S2 from RG7386, a Novel Tetravalent FAP-DR5 Antibody, Effectively Triggers FAP-Dependent, Avidity-Driven DR5 Hyperclustering and Tumor Cell Apoptosis
Authors: Brünker Peter , Wartha Katharina , Friess Thomas , Grau-Richards Sandra , Waldhauer Inja , Koller Claudia Ferrara , Weiser Barbara , Majety Meher , Runza Valeria , Niu Huifeng , Packman Kathryn , Feng Ningping , Daouti Sherif , Hosse Ralf J. , Mössner Ekkehard , Weber Thomas G. , Herting Frank , Scheuer Werner , Sade Hadassah , Shao Cuiying , Liu Bin , Wang Peng , Xu Gary , Vega-Harring Suzana , Klein Christian , Bosslet Klaus , Umaña Pablo
Published: April 2023

Figure S1. Related to Figure 1. Characterization of FAP-drozitumab BsAbs and antitumor efficacy in the presence of FAP-expressing fibroblasts in in vitro co-culture models; Table S1. BsAb binding to DR5 and FAP antigens; Table S2. Anti-tumor efficacy of FAP-drozitumab bispecific molecules is FAPdependent.
Supplementary Table S1: NGS-Cancer Panel from Genomic and Immunological Tumor Profiling Identifies Targetable Pathways and Extensive CD8+/PDL1+ Immune Infiltration in Inflammatory Breast Cancer Tumors
Authors: Hamm Christopher A. , Moran Diarmuid , Rao Kakuturu , Trusk Patricia B. , Pry Karen , Sausen Mark , Jones Siân , Velculescu Victor E. , Cristofanilli Massimo , Bacus Sarah
Published: April 2023

NGS-Cancer Panel Gene list
Figure S2 from Combined Inhibition of NEDD8-Activating Enzyme and mTOR Suppresses NF2 Loss–Driven Tumorigenesis
Authors: Cooper Jonathan , Xu Qingwen , Zhou Lu , Pavlovic Milica , Ojeda Virginia , Moulick Kamalika , de Stanchina Elisa , Poirier John T. , Zauderer Marjorie , Rudin Charles M. , Karajannis Matthias A. , Hanemann C. Oliver , Giancotti Filippo G.
Published: April 2023

Figure S2 related to Figure 2 shows that DCAF1 knockdown suppresses the growth of Merlin-mutant mesothelioma cells.
Fig S4 from CD3-Bispecific Antibody Therapy Turns Solid Tumors into Inflammatory Sites but Does Not Install Protective Memory
Authors: Benonisson Hreinn , Altıntaş Işıl , Sluijter Marjolein , Verploegen Sandra , Labrijn Aran F. , Schuurhuis Danita H. , Houtkamp Mischa A. , Verbeek J. Sjef , Schuurman Janine , van Hall Thorbald
Published: April 2023

In vivo T cell activation in tumor-free animals
Fig S3 from CD3-Bispecific Antibody Therapy Turns Solid Tumors into Inflammatory Sites but Does Not Install Protective Memory
Authors: Benonisson Hreinn , Altıntaş Işıl , Sluijter Marjolein , Verploegen Sandra , Labrijn Aran F. , Schuurhuis Danita H. , Houtkamp Mischa A. , Verbeek J. Sjef , Schuurman Janine , van Hall Thorbald
Published: April 2023

Depletion effiency and Tumor outgrowth curves
Table S2 from Preclinical Activity of Abemaciclib Alone or in Combination with Antimitotic and Targeted Therapies in Breast Cancer
Authors: O'Brien Neil , Conklin Dylan , Beckmann Richard , Luo Tong , Chau Kevin , Thomas Josh , Mc Nulty Ann , Marchal Christophe , Kalous Ondrej , von Euw Erika , Hurvitz Sara , Mockbee Colleen , Slamon Dennis J.
Published: April 2023

Abemaciclib response biomarkers, breast cancer cancer cell line panel
Supplementary Figure S2 from Preclinical Activity of Abemaciclib Alone or in Combination with Antimitotic and Targeted Therapies in Breast Cancer
Authors: O'Brien Neil , Conklin Dylan , Beckmann Richard , Luo Tong , Chau Kevin , Thomas Josh , Mc Nulty Ann , Marchal Christophe , Kalous Ondrej , von Euw Erika , Hurvitz Sara , Mockbee Colleen , Slamon Dennis J.
Published: April 2023

In vivo efficacy of abemaciclib plus hormonal blockade in ER+/HER2- breast cancer cell line xenografts
SupplementalFigures 1-4, Tables from Hypomorphic mTOR Downregulates CDK6 and Delays Thymic Pre-T LBL Tumorigenesis
Authors: Gary Joy M. , Simmons John K. , Xu Jinfei , Zhang Shuling , Peat Tyler J. , Watson Nicholas , Gamache Benjamin J. , Zhang Ke , Kovalchuk Alexander L. , Michalowski Aleksandra M. , Chen Jin-Qiu , Thaiwong Tuddow , Kiupel Matti , Gaikwad Snehal , Etienne Maudeline , Simpson R. Mark , Dubois Wendy , Testa Joseph R. , Mock Beverly A.
Published: April 2023

Supplemental Figures S1-S4 + two supp. tables
Table S2 from Development of AO-176, a Next-Generation Humanized Anti-CD47 Antibody with Novel Anticancer Properties and Negligible Red Blood Cell Binding
Authors: Puro Robyn J. , Bouchlaka Myriam N. , Hiebsch Ronald R. , Capoccia Benjamin J. , Donio Michael J. , Manning Pamela T. , Frazier William A. , Karr Robert W. , Pereira Daniel S.
Published: April 2023

Table S2 is a summary of AO-176 tumor cell killing in solid and hematologic tumor cell lines
Figure S1 from Development of AO-176, a Next-Generation Humanized Anti-CD47 Antibody with Novel Anticancer Properties and Negligible Red Blood Cell Binding
Authors: Puro Robyn J. , Bouchlaka Myriam N. , Hiebsch Ronald R. , Capoccia Benjamin J. , Donio Michael J. , Manning Pamela T. , Frazier William A. , Karr Robert W. , Pereira Daniel S.
Published: April 2023

Figure S1 depicts the sequence and analytical characteristics of AO-176
Figure S3 from Site-Specific PEGylation of Anti-Mesothelin Recombinant Immunotoxins Increases Half-life and Antitumor Activity
Authors: Zheng Zeliang , Okada Ryuhei , Kobayashi Hisataka , Nagaya Tadanobu , Wei Junxia , Zhou Qi , Lee Fred , Bera Tapan K. , Gao Yun , Kuhlman William , Tai Chin-Hsien , Pastan Ira
Published: April 2023

ADP ribosylation activities of PEGylated RITs
Figure S3 from An Orally Available Tubulin Inhibitor, VERU-111, Suppresses Triple-Negative Breast Cancer Tumor Growth and Metastasis and Bypasses Taxane Resistance
Authors: Deng Shanshan , Krutilina Raisa I. , Wang Qinghui , Lin Zongtao , Parke Deanna N. , Playa Hilaire C. , Chen Hao , Miller Duane D. , Seagroves Tiffany N. , Li Wei
Published: April 2023

Figure S3 shows that the IC50 dose of VERU-111 inhibits wound healing of TNBC cells in a scratch wound healing assay. The scratch assay was repeated in MDA-MB-231 and MDA-MB-468 cells at 8 nM concentration. (A) Representative images of MDA-MB-231 cells treated with 8 nM of VERU-111 at 12h and 24h post-scratch as compared to untreated cells. (B) Representative images of MDA-MB-468 cells treated with 8 nM of VERU-111 at 48h and 96h post-scratch as compared to untreated cells. Blue lines indicate the wound edges and the bar graphs show the percent relative wound density (wound fill-in) at each time point. (C) The concentration-dependent effect of VERU-111 on MDA-MB-231 cell wound healing over time was measured following wounding with the WoundMaker tool using the IncuCyte S3 live cell imager. Data were collected every 2 h and analyzed using the cell migration algorithm. Exposure to 8, 12 or 16 nM of VERU-111 inhibits the migration of MDA-MB-231 cells in a concentration-dependent manner. All data are presented as the grand mean {plus minus} SEM of three independent experiments.
Table S5 from Identification of Predictive ERBB Mutations by Leveraging Publicly Available Cell Line Databases
Authors: Koivu Marika K.A. , Chakroborty Deepankar , Tamirat Mahlet Z. , Johnson Mark S. , Kurppa Kari J. , Elenius Klaus
Published: April 2023

Table S5 shows the cBioPortal prediction of potential clinical implication of ERBB mutations.
Figure S5 from Design of a Cytotoxic Neuroblastoma-Targeting Agent Using an Enzyme Acting on Polysialic Acid Fused to a Toxin
Authors: Lehti Timo A. , Pajunen Maria I. , Jokilammi Anne , Korja Miikka , Lilie Hauke , Vettenranta Kim , Finne Jukka
Published: April 2023

Figure S5 shows the apoptotic effects of DT386F-endoNA2 on neuroblastoma cells.
Figure S3 from Design of a Cytotoxic Neuroblastoma-Targeting Agent Using an Enzyme Acting on Polysialic Acid Fused to a Toxin
Authors: Lehti Timo A. , Pajunen Maria I. , Jokilammi Anne , Korja Miikka , Lilie Hauke , Vettenranta Kim , Finne Jukka
Published: April 2023

Figure S3 shows the effect of S706A mutation on binding and internalization activities of endoNA2.
Supplementary Data from A Precision Medicine Drug Discovery Pipeline Identifies Combined CDK2 and 9 Inhibition as a Novel Therapeutic Strategy in Colorectal Cancer
Authors: Somarelli Jason A. , Roghani Roham Salman , Moghaddam Ali Sanjari , Thomas Beatrice C. , Rupprecht Gabrielle , Ware Kathryn E. , Altunel Erdem , Mantyh John B. , Kim So Young , McCall Shannon J. , Shen Xiling , Mantyh Christopher R. , Hsu David S.
Published: April 2023

Supplementary Figures
Supplementary Data Summary from Development of a MUC16-Targeted Near-Infrared Fluorescent Antibody Conjugate for Intraoperative Imaging of Pancreatic Cancer
Authors: Olson Madeline T. , Wojtynek Nicholas E. , Talmon Geoffrey A. , Caffrey Thomas C. , Radhakrishnan Prakash , Ly Quan P. , Hollingsworth Michael A. , Mohs Aaron M.
Published: April 2023

Compilation of supplemental figures.
Supplementary Methods, Supplementary Tables 1 - 11, and Supplementary Figures 1-9 from Characterization of a Novel FLT3 BiTE Molecule for the Treatment of Acute Myeloid Leukemia
Authors: Brauchle Bettina , Goldstein Rebecca L. , Karbowski Christine M. , Henn Anja , Li Chi-Ming , Bücklein Veit L. , Krupka Christina , Boyle Michael C. , Koppikar Priya , Haubner Sascha , Wahl Joachim , Dahlhoff Christoph , Raum Tobias , Rardin Matthew J. , Sastri Christine , Rock Dan A. , von Bergwelt-Baildon Michael , Frank Brendon , Metzeler Klaus H. , Case Ryan , Friedrich Matthias , Balazs Mercedesz , Spiekermann Karsten , Coxon Angela , Subklewe Marion , Arvedson Tara
Published: April 2023

Supplementary Methods. Supplementary Table 1 - Key Resources Including Sources of Biological Samples, Antibodies, and Key Reagents. Supplementary Table 2 -Correlation of FLT3 Expression Levels With Disease Characteristics at Initial Diagnosis. Supplementary Table 3 - Summary of Human Nonimmune Tissues Positive for FLT3 mRNA and/or Protein Expression in At Least One Platform. Supplementary Table 4. Cerebellum FLT3 RNA Analysis. Supplementary Table 5 - Binding Affinities of Experimental FLT3 BiTE® Molecule and AMG 427. Supplementary Table 6 - Cell Line Characterization, Cytotoxicity, and T-Cell Activation (Human). Supplementary Table 7 - Cynomolgus Monkey Pharmacokinetics for Experimental FLT3 BiTE® Molecule. Supplementary Table 8 - Cynomolgus Monkey Pharmacokinetics for AMG 427. Supplementary Table 9 - Cynomolgus Monkey FLT3 Primer and Probe Sequences. Supplementary Table 10 - Patient Characteristics. Supplementary Table 11 - Mass Spectrometry Results. Supplementary Figure 1 - Additional flow cytometry expression analysis for acute myeloid leukemia (AML) and healthy donor (HD) bone marrow (BM). Supplementary Figure 2 - Analysis of FLT3 expression on normal human bone marrow. Supplementary Figure 3 - Analysis of FLT3 transcript and protein in cerebellum revealed truncated transcripts. Supplementary Figure 4 - In vitro characterization of FLT3 BiTE® molecules. Supplementary Figure 5 - Experimental FLT3 BiTE® molecule activity in vivo. Supplementary Figure 6 - FLT3 BiTE® molecule in vitro activity with cynomolgus monkey effector cells. Supplementary Figure 7 - FLT3 expression in cynomolgus monkey blood and bone marrow. Supplementary Figure 8 - AMG 427 pharmacodynamics in cynomolgus monkey. Supplementary Figure 9 - (A) Representative populations of T cells (CD2+) and AML cells (CD33+) over time from a responder (Patient 4; Figure 6A). (B) PD-1 expression on CD3+ cells from a T-cell-dependent cellular cytotoxicity (TDCC) of human pan T cells co-cultured 5:1 with MV4-11 cells for 48 hours assessed by flow cytometry.
Supplementary Figure 3 from Novel Humanized Mesothelin-Expressing Genetically Engineered Mouse Models Underscore Challenges in Delivery of Complex Therapeutics to Pancreatic Cancers
Authors: Hagerty Brendan , O'Sullivan T. Norene , Zhang Xianyu , Collins N. Keith , Custer Lawrence Wendi , Bassel Laura L. , Pate Nathan , Xu Jian , Guerin Theresa M. , Kozlov Serguei , Alewine Christine
Published: April 2023

Supplemental Figure 3. A) Immunostaining with anti-hMSLN antibody in orthotopic KPCdelMsln+hMSLN tumors following LMB-100 treatment. Scale bar = 800 Âµm. B) IP and orthotopic KPCdelMsln+hMSLN tumors were stained with Maisson Trichrome and percent positive pixels was quantified by image analysis. Each point represents quantification from a separate tumor/animal.
Figure S2 from The Indenoisoquinoline LMP517: A Novel Antitumor Agent Targeting both TOP1 and TOP2
Authors: Marzi Laetitia , Sun Yilun , Huang Shar-yin N. , James Amy , Difilippantonio Simone , Pommier Yves
Published: April 2023

Cellular DNA damage induced at nanomar concentrations of LMP517
Supplementary Data from Novel Auristatins with High Bystander and Cytotoxic Activities in Drug Efflux–positive Tumor Models
Authors: Moquist Philip N. , Bovee Tim D. , Waight Andrew B. , Mitchell Jamie A. , Miyamoto Jamie B. , Mason Marsha L. , Emmerton Kim K. , Stevens Nicole , Balasubramanian Cindy , Simmons Jessica K. , Lyon Robert P. , Senter Peter D. , Doronina Svetlana O.
Published: April 2023

Supplementary Data
Supplementary Figure 2 from The Role of Specific ATP-Binding Cassette Transporters in the Acquired Resistance to Pyrrolobenzodiazepine Dimer–Containing Antibody–Drug Conjugates
Authors: Corbett Simon , Huang Shiran , Zammarchi Francesca , Howard Philip W. , van Berkel Patrick H. , Hartley John A.
Published: April 2023

Supplementary Figure 2. Qiagen RT2 PCR array of human DNA damage response pathway genes on Karpas-299 and NCI-N87 ADC and PBD resistant cell lines. A. Volcano plot of DDR gene expression relative to wt in Karpas ADCr cell line. B. Volcano plot of DDR gene expression relative to wt in Karpas PBDr cell line. C. Volcano plot of DDR gene expression relative to wt in NCI ADCr cell line. D. Volcano plot of DDR gene expression relative to wt in NCI PBDr cell line.
Supplementary Methods from MMAE Delivery Using the Bicycle Toxin Conjugate BT5528
Authors: Bennett Gavin , Brown Amy , Mudd Gemma , Huxley Philip , Van Rietschoten Katerine , Pavan Silvia , Chen Liuhong , Watcham Sophie , Lahdenranta Johanna , Keen Nicholas
Published: April 2023

Supplementary information detailing methods used to generate in vitro and in vivo data presented in paper
Figure S3 from BRCA1/MAD2L1 Deficiency Disrupts the Spindle Assembly Checkpoint to Confer Vinorelbine Resistance in Mesothelioma
Authors: Busacca Sara , O'Regan Laura , Singh Anita , Sharkey Annabel J. , Dawson Alan G. , Dzialo Joanna , Parsons Aimee , Kumar Neelam , Schunselaar Laurel M. , Guppy Naomi , Nakas Apostolos , Sheaff Michael , Mansfield Aaron S. , Janes Sam M. , Baas Paul , Fry Andrew M. , Fennell Dean A.
Published: April 2023

ROC curves for BRCA1 and MAD2L1 for explants. The percentage of cleaved PARP was used as a measure of outcome
Supplementary Methods, Supplementary tables 1-3 and figures 1-3 from FGFR Inhibition Enhances Sensitivity to Radiation in Non–Small Cell Lung Cancer
Authors: SenthilKumar Gopika , Fisher Michael M. , Skiba Justin H. , Miller Margot C. , Brennan Sean R. , Kaushik Saakshi , Bradley Samantha T. , Longhurst Colin A. , Buehler Darya , Nickel Kwangok P. , Iyer Gopal , Kimple Randall J. , Baschnagel Andrew M.
Published: April 2023

Supplementary Table 1. Short Tandem Repeat analysis of cell lines used in this study. Supplementary Table 2. Antibodies used in this study. Supplemental Table 3: Dose Enhancement factors at survival fraction of 0.1 (DEF0.1) and Pvalues for radiation clonogenic curves. Western blot showing FGFR1, FGFR2 and FGFR3 protein levels in HTE and BeasB2 cell lines. Supplementary figure 2. Normalized intensity quantification of pMAPK and pAKT proteins determined by western blot after treatment with AZD4547 in three NSCLC cell lines. Supplementary figure 3. Combined radiation and AZD4547 induces autophagy.
Table S1 from STimulator of INterferon Genes Agonism Accelerates Antitumor Activity in Poorly Immunogenic Tumors
Authors: Perera Samanthi A. , Kopinja Johnny E. , Ma Yanhong , Muise Eric S. , Laskey Jason , Chakravarthy Kalyan , Chen Yiping , Cui Long , Presland Jeremy , Sathe Manjiri , Javaid Sarah , Minnihan Ellen C. , Ferguson Heidi M. , Piesvaux Jennifer , Pan Bo-Sheng , Zhao Shuxia , Sharma Sharad K. , Woo Hyun Chong , Pucci Vincenzo , Knemeyer Ian , Cemerski Saso , Cumming Jared , Trotter B. Wesley , Tse Archie , Khilnani Anuradha , Ranganath Sheila , Long Brian J. , Bennett David Jonathan , Addona George H.
Published: April 2023

Supplemental Table 1: Log2Ratio and p values for all 310 genes shown in Figure 6
Table S1 from Gene Expression Signature Correlates with Outcomes in Metastatic Renal Cell Carcinoma Patients Treated with Everolimus Alone or with a Vascular Disrupting Agent
Authors: Yang Eddy S. , Nassar Amin H. , Adib Elio , Jegede Opeyemi A. , Alaiwi Sarah Abou , Manna Deborah L. Della , Braun David A. , Zarei Mahsa , Du Heng , Pal Sumanta K. , Naik Gurudatta , Sonpavde Guru P.
Published: April 2023

Master table of 82 mRCC patients with clinical and transcriptomic Nanostring data
Supplementary Data from Gene Expression Signature Correlates with Outcomes in Metastatic Renal Cell Carcinoma Patients Treated with Everolimus Alone or with a Vascular Disrupting Agent
Authors: Yang Eddy S. , Nassar Amin H. , Adib Elio , Jegede Opeyemi A. , Alaiwi Sarah Abou , Manna Deborah L. Della , Braun David A. , Zarei Mahsa , Du Heng , Pal Sumanta K. , Naik Gurudatta , Sonpavde Guru P.
Published: April 2023

Supplemtary figure and table legends
Dolaflexin_Figure1 from The Dolaflexin-based Antibody–Drug Conjugate XMT-1536 Targets the Solid Tumor Lineage Antigen SLC34A2/NaPi2b
Authors: Bodyak Natalya D. , Mosher Rebecca , Yurkovetskiy Aleksandr V. , Yin Mao , Bu Charlie , Conlon Patrick R. , Demady Damon R. , DeVit Michael J. , Gumerov Dmitry R. , Gurijala Venu R. , Lee Winnie , McGillicuddy Dennis , Park Peter U. , Poling Laura L. , Protopova Marina , Qin LiuLiang , Stevenson Cheri A. , Ter-Ovanesyan Elena , Uttard Alex , Xiao Dongmei , Xu Jian , Xu Ling , Bergstrom Donald A. , Lowinger Timothy B.
Published: April 2023

These are figures associated with a second manuscript that has been submitted for consideration for co-publication. It describes the platform and hopefully will facilitate the reviewers' assessment of this manuscript on XMT-1536. we are also happy to make this available as supplementary data for this publication if desired.
Supplementary Figure S3 from NAP1051, a Lipoxin A4 Biomimetic Analogue, Demonstrates Antitumor Activity Against the Tumor Microenvironment
Authors: Dong Tiange , Dave Priyal , Yoo EunJeong , Ebright Brandon , Ahluwalia Kabir , Zhou Eugene , Asante Isaac , Salimova Malika , Pei Hua , Lin Tracey , Mead Andrew , Li Zeyang , Humayun Mark , Petasis Nicos A. , Epstein Alan L. , Louie Stan G.
Published: April 2023

Figure S3 shows the gating strategy for analysis of immune cell subpopulations in the spleen and tumor.
Supplementary Figure S1 from NAP1051, a Lipoxin A4 Biomimetic Analogue, Demonstrates Antitumor Activity Against the Tumor Microenvironment
Authors: Dong Tiange , Dave Priyal , Yoo EunJeong , Ebright Brandon , Ahluwalia Kabir , Zhou Eugene , Asante Isaac , Salimova Malika , Pei Hua , Lin Tracey , Mead Andrew , Li Zeyang , Humayun Mark , Petasis Nicos A. , Epstein Alan L. , Louie Stan G.
Published: April 2023

Figure S1 shows the gating strategy for PMN apoptosis analysis, RT-PCR, immunofluorescence staining, and flow cytometry analyses of FPR2/ALX-transfected HEK cells, effect of NAP1051 on dTHP-1 cells with selective kinase inhibitors, and Western blot with normal HEK cells and FPR2/ALX-transfected HEK cells
Supplementary Materials and Methods from Preclinical Studies of OBI-999: A Novel Globo H–Targeting Antibody–Drug Conjugate
Authors: Yang Ming-Chen , Shia Chi-Sheng , Li Wan-Fen , Wang Chun-Chung , Chen I-Ju , Huang Teng-Yi , Chen Yu-Jung , Chang Hui-Wen , Lu Chi-Huan , Wu Yueh-Chin , Wang Nan-Hsuan , Lai Jiann-Shiun , Yu Cheng-Der , Lai Ming-Tain
Published: April 2023

Supplementary Materials and Methods
Supplementary Table and Figures from Resistance Profile and Structural Modeling of Next-Generation ROS1 Tyrosine Kinase Inhibitors
Authors: Keddy Clare , Shinde Pushkar , Jones Kristen , Kaech Stefanie , Somwar Romel , Shinde Ujwal , Davare Monika A.
Published: April 2023

Supplementary Table and Figures
Supplementary Table and Figures
Supplementary Figure from Combination of Ribociclib and Gemcitabine for the Treatment of Medulloblastoma
Authors: Pribnow Allison , Jonchere Barbara , Liu Jingjing , Smith Kyle S. , Campagne Olivia , Xu Ke , Robinson Sarah , Patel Yogesh , Onar-Thomas Arzu , Wu Gang , Stewart Clinton F. , Northcott Paul A. , Yu Jiyang , Robinson Giles W. , Roussel Martine F.
Published: April 2023

Supplementary Figure from Combination of Ribociclib and Gemcitabine for the Treatment of Medulloblastoma
Supplementary Materials, Methods, and References SM1 from Sensitization to Ionizing Radiation by MEK Inhibition Is Dependent on SNAI2 in Fusion-Negative Rhabdomyosarcoma
Authors: Hensch Nicole R. , Bondra Kathryn , Wang Long , Sreenivas Prethish , Zhao Xiang R. , Modi Paulomi , Vaseva Angelina V. , Houghton Peter J. , Ignatius Myron S.
Published: April 2023

Supplementary Materials, Methods, and References
Supplementary Figures S1-S7 from ER-851, a Novel Selective Inhibitor of AXL, Overcomes Resistance to Antimitotic Drugs
Authors: Tsukamoto Shuntaro , Sugi Naoko Hata , Nishibata Kyoko , Nakazawa Youya , Ito Daisuke , Fukushima Sayo , Nakagawa Takayuki , Ichikawa Kenji , Kato Yu , Kakiuchi Dai , Goto Aya , Itoh-Yagi Machiko , Aota Tomoki , Inoue Satoshi , Yamane Yoshinobu , Murai Norio , Azuma Hiroshi , Nagao Satoshi , Sasai Ken , Akagi Tsuyoshi , Imai Toshio , Matsui Junji , Matsushima Tomohiro
Published: April 2023

Supplementary Data Figure S1-S7
Supplementary Data from Optimization and Characterization of a Bone Culture Model to Study Prostate Cancer Bone Metastasis
Authors: Wu Yi-Hsuan , Gugala Zbigniew , Barry Megan M. , Shen Yichao , Dasgupta Subhamoy , Wang Hai
Published: April 2023

Supplementary Data from Optimization and Characterization of a Bone Culture Model to Study Prostate Cancer Bone Metastasis
Supplementary Figure from NOTCH Signaling Limits the Response of Low-Grade Serous Ovarian Cancers to MEK Inhibition
Authors: Llaurado Fernandez Marta , Hijmans E. Marielle , Gennissen Annemiek M.C. , Wong Nelson K.Y. , Li Shang , Wisman G. Bea A. , Hamilton Aleksandra , Hoenisch Joshua , Dawson Amy , Lee Cheng-Han , Bittner Madison , Kim Hannah , DiMattia Gabriel E. , Lok Christianne A.R. , Lieftink Cor , Beijersbergen Roderick L. , de Jong Steven , Carey Mark S. , Bernards René , Berns Katrien
Published: April 2023

Supplementary Figure from NOTCH Signaling Limits the Response of Low-Grade Serous Ovarian Cancers to MEK Inhibition
Supplementary Data from FGFR4-Targeted Chimeric Antigen Receptors Combined with Anti-Myeloid Polypharmacy Effectively Treat Orthotopic Rhabdomyosarcoma
Authors: Sullivan Peter M. , Kumar Rajesh , Li Wei , Hoglund Virginia , Wang Lingyan , Zhang Yue , Shi Megan , Beak Dusan , Cheuk Adam , Jensen Michael C. , Khan Javed , Dimitrov Dimiter S. , Orentas Rimas J.
Published: April 2023

Supplementary Data from FGFR4-Targeted Chimeric Antigen Receptors Combined with Anti-Myeloid Polypharmacy Effectively Treat Orthotopic Rhabdomyosarcoma
Supplementary Table from PH-Binding Motif in PAR4 Oncogene: From Molecular Mechanism to Drug Design
Authors: Nag Jeetendra Kumar , Malka Hodaya , Sedley Shoshana , Appasamy Priyanga , Rudina Tatyana , Levi Tgst , Hoffman Amnon , Gilon Chaim , Uziely Beatrice , Bar-Shavit Rachel
Published: April 2023

Supplementary Table from PH-Binding Motif in PAR4 Oncogene: From Molecular Mechanism to Drug Design
Abstract 127: Single-cell methylation sequencing data reveals succinct metastatic migration histories and tumor progression models
Authors: Li Xuan C. , Liu Yuelin , Mehrabadi Farid Rashidi , Schäffer Alejandro A. , Pratt Drew , Crawford David R. , Malikić Salem , Molloy Erin K. , Gopalan Vishaka , Mount Stephen M. , Ruppin Eytan , Aldape Kenneth , Sahinalp S. Cenk
Published: April 2023

Recent studies exploring the impact of methylation in tumor evolution suggest that while the methylation status of many of the CpG sites are preserved across distinct lineages, others are altered as the cancer progresses. Since changes in methylation status of a CpG site may be retained in mitosis, they could be used to infer the progression history of a tumor via single-cell lineage tree reconstruction. In this work, we introduce the first principled distance-based computational method, Sgootr, for inferring a tumor's single-cell methylation lineage tree and jointly identifying lineage-informative CpG sites which harbor changes in methylation status that are retained along the lineage. We apply Sgootr on the single-cell bisulfite-treated whole genome sequencing data of multiregionally-sampled tumor cells from 9 metastatic colorectal cancer patients made available by Bian et al., as well as multiregionally-sampled single-cell reduced-representation bisulfite sequencing data from a glioblastoma patient made available by Chaligne et al. We demonstrate that the tumor lineages constructed reveal a simple model underlying colorectal tumor progression and metastatic seeding. A comparison of Sgootr against alternative approaches shows that Sgootr can construct lineage trees with fewer migration events and more in concordance with the sequential-progression model of tumor evolution, in time a fraction of that used in prior studies. Interestingly, lineage-informative CpG sites identified by Sgootr are in inter-CpG island (CGI) regions, as opposed to CGI's, which have been the main regions of interest in genomic methylation-related analyses. Sgootr is implemented as a Snakemake workflow, available at https://github.com/liuy0421/Sgootr.

Citation Format: Xuan C. Li, Yuelin Liu, Farid Rashidi Mehrabadi, Alejandro A. Schäffer, Drew Pratt, David R. Crawford, Salem Malikić, Erin K. Molloy, Vishaka Gopalan, Stephen M. Mount, Eytan Ruppin, Kenneth Aldape, S. Cenk Sahinalp. Single-cell methylation sequencing data reveals succinct metastatic migration histories and tumor progression models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 127.
Abstract 1274: Extrinsic photoaging vs intrinsic aging in dermal fibroblasts: Its impact on the microenvironment and melanoma progression
Authors: Wang Vania , Chhabra Yash , Hueser Laura , Pramod Sneha , Carey Alexis , Weeraratna Ashani
Published: April 2023

Melanoma is the deadliest form of skin cancer, with its incidence rising disproportionally to other cancers. Chronological aging is a major risk factor in all types of cancer as it has been shown to drive changes in noncancerous cells (stromal and immune) of the tumor microenvironment (TME) to promote tumor initiation and progression. Ultraviolet radiation (UVR) exposure, which is a major risk factor for melanoma, also can cause a physical premature aging phenotype, known as photoaging. Photoaging of the skin often occurs in tandem with chronological aging, and contributes to the physical features of aging through similar alterations in normal skin structure (eg. collagen degradation). However, the molecular mechanisms of how photoaging compares to chronological aging in changing the TME and on tumor progression is still lacking.Preliminary data have indicated that photoaging of young dermal fibroblasts induce some similar signature changes to that of intrinsically aged. We have shown in-vitro that UVA exposure in young fibroblasts cause upregulation in MMP-1, a matrix remodeling protein, and downregulation in β-catenin, the cell adhesion transcription regulator. These same changes were also shown in chronologically aged fibroblasts. In addition, both photoaging and chronological aging decreases ECM structural integrity, with increased collagen fragmentation and a reduction in the renewal of cross-linking proteins. To investigate how photoaging contributes to the pro-tumorigenic microenvironment of the aged dermis, we are using biochemical and phenotypic assays to study how chronic UVA exposure induces an intrinsically aged-like fibroblast behavior and secretome. We also hypothesize that photoaged fibroblasts reprogram the TME to become more permissible for the growth of tumor cells, and will test changes in invasion and metastasis or melanoma both in-vitro and in-vivo.Studying the impact of photoaging on aging in the skin microenvironment will help identify therapeutic targets and have direct clinical applications for cancer patients.

Citation Format: Vania Wang, Yash Chhabra, Laura Hueser, Sneha Pramod, Alexis Carey, Ashani Weeraratna. Extrinsic photoaging vs intrinsic aging in dermal fibroblasts: Its impact on the microenvironment and melanoma progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1274.
Abstract 1364: Profiling tumor microenvironment for therapeutic intervention to soft-tissue sarcomas
Authors: Xiao Jin-Fen , Broz Marina , Piras Roberta , Ishaya Kristin , Ko Emily , Guarnerio Jlenia
Published: April 2023

Soft-tissue sarcomas (STS) are mesenchymal tumors having remarkably diverse histological features. Genomic studies reported that STS have low frequencies of genetic mutation, but often harboring copy number variations (CNVs). For STS, CNVs frequently happen in essential tumor suppressor genes and oncogenic transcriptional genes, neither of which are easily “druggable” targets. Moreover, in concordance with low tumor mutation burden rate, STS show low immunogenicity with less cytotoxic TIL in the TME and is unfortunately linked to a lower responsive rate to PD1/PD-L1 ICB. Using a syngeneic mouse UPS model, we previously found that tumor-associated macrophages (TAM) in the TME are promoting the growth of sarcoma. We further identified specific tumor microenvironmental elements contributing to the pro-tumorigenicity of TAMs, which represent a promising novel clinical target for myeloid-related intervention. Therefore, we hypothesize that exploration of TME is beneficial to identify clinically promising targets for STS. To study the TME in STS, we performed bulk RNA-seq and scRNA-seq on human sarcoma samples which covered different histological subtypes of STS and compare them to normal samples. Our results show that, unlike carcinomas originating from epithelial cells, extracellular matrix (ECM)-related genes are highly upregulated in both cancer-associated fibroblast (CAF) and sarcoma tumor cells. Further analysis show that upregulation of these ECM genes is associated with a worse survival outcome in sarcoma patients. In order to further explore the STS and TME orchestra, we established several syngeneic mouse models according to the genetic aberrations reported in TCGA data. We are currently characterizing these syngeneic mouse models to find out whether they recapitulate human STS samples. Our goal is to utilize these mouse models to study the relationship of ECM-related genes in both CAF and tumor cells, find out the effect of upregulated ECM-related genes on the immune compartment in TME and search for potential therapeutic interventions.

Citation Format: Jin-Fen Xiao, Marina Broz, Roberta Piras, Kristin Ishaya, Emily Ko, Jlenia Guarnerio. Profiling tumor microenvironment for therapeutic intervention to soft-tissue sarcomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1364.
Abstract 1457: PDEF restricts prostate cancer cellular plasticity in part by modulating YAP1/TAZ-TEAD transcriptional network
Authors: Koul Hari K. , Jaiswal Praveen K. , Mohajan Suman , Vatanmakanian Mousa , Wang Fengtian , Koul Sweaty
Published: April 2023

Background: The Hippo TEAD-transcriptional regulators YAP1 and TAZ play essential role in cancer cell growth and metastasis. However, the function of YAP1 and TAZ in prostate cancer is not well characterized. We discovered that expression of PDEF is decreased during prostate cancer progression and re-expression of PDEF limits prostate cancer metastasis in part by promoting luminal epithelial phenotype, but mechanisms of PDEF action are not completely understood. In the present study, we evaluated the effects of the SPDEF on YAP1/TAZ levels and expression of YAP1/TAZ regulated genes in prostate cancer (PCa) cells. We also evaluated expression of SPDEF, YAP1, TAZ and YAPTAZZ-Tead regulated genes in clinical cohorts of prostate cancer patients.

Material & Method: Prostate cancer (PC3 and DU145) cells were transfected with PDEF or respective vector control. shRNA was used to knock down SPDEF in AR positive and androgen responsive LNCaP cells. Protein levels were analyzed by western blotting. Gene expression was monitored by microarray analysis/RNA seq and confirmed by RTPCR assays. Cell migration and invasion were measured by scratch wound healing and by trans well migration though Matrigel assays respectively. We analyzed publicly available gene expression data in several prostate cancer cohorts.

Results: We observed that the two well-characterized metastatic PCa cells (PC3 and DU145 cells) express both YAP1 and TAZ, but do not express SPDEF. Expression of PDEF in PC3 and DU145 cells resulted in an increased phospho-YAP1 and phospho-TAZ protein levels and inhibition of YAP1/TAZ target genes, as compared to the respective vector control, directly demonstrating that PDEF plays a critical role in modulating YAP1/TAZ-TEAD transcriptional activity, and by extension in the regulation of the Hippo pathway. Analysis of publicly available PCa data sets revealed a significant increase in TAZ mRNA levels in prostate cancer tissues as compared to normal prostate tissues (p=4.95E-08). Furthermore, we also found a significant gradual increase in TAZ mRNA expression and concomitant decrease in YAP1 mRNA during disease progression and metastasis and in neuroendocrine PCa.

Conclusions: Taken together these data suggest that SPDEF limits prostate cancer metastasis in part by targeting YAP1/TAZ driven transcriptional output.

Citation Format: Hari K. Koul, Praveen K. Jaiswal, Suman Mohajan, Mousa Vatanmakanian, Fengtian Wang, Sweaty Koul. PDEF restricts prostate cancer cellular plasticity in part by modulating YAP1/TAZ-TEAD transcriptional network [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1457.
Abstract 1529: Characterizing antibody internalization for rational selection of ADC linker design
Authors: Shao Zekun , Panyam Jayanth
Published: April 2023

Antibody-drug conjugates (ADC), which target a highly potent cytotoxin to a specific protein overexpressed on tumor cells, are a promising class of anticancer therapeutics. In addition to the antibody characteristics, intracellular reactivity of the antibody-cytotoxin linker (which affects drug release) is a key determinant of ADC efficacy. While many different linkers have been used in approved ADCs, the intracellular trafficking profile of an antibody will likely impact the effectiveness of different linkers differently. Here, we determined the cellular internalization of two antibodies, anti-perlecan antibody (‘AM6’), and anti-EGFR antibody (cetuximab; ‘Ctx’), in MDA-MB-231-LM2 cells (a lung-metastatic TNBC cell line). Using live-cell confocal microscopy, we investigated the effect of endocytosis inhibitors on intracellular trafficking of AM6 and Ctx. While AM6 was internalized quicker compared to Ctx, the internalization of both antibodies was reduced significantly by 5-(N-Ethyl-N-isopropyl)-amiloride, a macropinocytosis inhibitor and, to a lesser extent, by chlorpromazine, an inhibitor of clathrin-mediated-endocytosis. The results suggest both AM6 and Ctx were internalized by both macropinocytosis and clathrin-mediated endocytosis. We then investigated whether acidic pH in the lysosomes and the reductive environment in the endosomal vesicles would facilitate drug release from ADC. We conjugated thiol-reactive doxorubicin to reduced interchain disulfides of Ctx via either the acid and glutathione (GSH)-labile pyridinyldithio-hydrazide linker or the non-cleavable C6-maleimide linker. Intracellular doxorubicin release for the two ADCs was evaluated by live-cell confocal microscopy. Doxorubicin was successfully conjugated to Ctx, with a DAR of 4.8 and 8.6 for the cleavable and non-cleavable linkers, respectively. Conjugation of doxorubicin to the antibody was confirmed by fluorescent gel imaging after SDS-PAGE and by using hydrophobic interaction chromatography. Confocal microscopic studies showed that the ADC with cleavable linker resulted in greater doxorubicin accumulation in the nucleus compared to the ADC with non-cleavable linker, which was predominantly sequestered in the lysosomes. These results suggest that the cleavable linker results in better drug release compared to the non-cleavable linker for the Ctx-doxorubicin ADC. Studies examining the effect of linker chemistry on drug release from the anti-perlecan ADC and on the anticancer efficacy of the two ADCs are ongoing.

Citation Format: Zekun Shao, Jayanth Panyam. Characterizing antibody internalization for rational selection of ADC linker design [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1529.