bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2538: MiR-92b targets FBXW7 in GBM cells
Authors: Sanchez-Alvarez Annelis Odette , Grafals-Ruiz Nilmary , Santana-Rivera Yasmarie , Valiyeva Fatma , Rosado-Rivera Nathaly M. , Vivas-Mejía Pablo E.
Published: April 2023

MicroRNA (miRNA) dysregulation plays a central role in the initiation, progression, and drug resistance of virtually all cancer types. MiRNA-based therapies are a promising therapeutic modality against most malignancies. MiR-92b is aberrantly abundant in glioblastoma (GBM), the most aggressive and mortal of all primary brain tumors. Thus, miR-92b is a potential target for GBM therapy. First, it is critically important to assess the molecular mechanism of miR-92b regulation in GBM cells. In this study, we investigated the molecular and biological effects of targeting miR-92b in GBM cells with oligonucleotide miRNA inhibitors (OMIs). Incubation of GBM cells with miR-92b-OMIs resulted in decreased cell proliferation and reduction in cell migration. MiR-92b inhibition also promoted apoptosis as assessed by caspase-3 activation. Bioinformatics and real-time PCR studies identified several miR-92b target genes in GBM cells. Western blot analysis and luciferase reporter assays confirmed F-Box and WD Repeat Domain Containing 7 (FBXW7) as a direct miR-92b target gene in GBM cells. FBXW7 is one of ~70 F-BOX proteins in humans particularly studied in cancer due to its role in the ubiquitylation of positive regulators; also known as accelerators of the cell cycle (i.e. the oncogene c-Myc). Thus, an increase in FBXW7 promoted the decrease of c-MYC protein levels in GBM cells. These findings indicate that targeting miR-92b inhibits GBM cell proliferation by increasing the levels of tumor suppressor genes in charge of protein ubiquitination.

Citation Format: Annelis Odette Sanchez-Alvarez, Nilmary Grafals-Ruiz, Yasmarie Santana-Rivera, Fatma Valiyeva, Nathaly M. Rosado-Rivera, Pablo E. Vivas-Mejía. MiR-92b targets FBXW7 in GBM cells [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 2538.
Abstract 2550: Growth factor-induced cell membrane macroplatforms drive protein kinase signaling for transformed cell growth
Authors: Alberti Saverio , Relli Valeria , Tripaldi Romina , Simeone Pasquale , Guerra Emanuela , Sacchetti Andrea , Ciufici Paolo , Moschella Antonino , Caiolfa Valeria , Zamai Moreno , Trerotola Marco
Published: April 2023

Nanoscopic, millisecond-lasting membrane platforms are modeled to constitute main functional signaling units. However, multiple signaling events in cell growth induction, including receptor Tyr-kinase (RTK) stimulation, calcium signaling, cytoplasmic kinase recruitment, operate over much broader space-time frames. Through dynamic confocal/super-resolution/electron-microscopy on living cells, we discovered that signaling competence is acquired through growth factor-induced coalescence of activated growth inducers (RTK, CD9, CD81, CD98, Co-029, CD316, Trop-1, Trop-2, PI3K, PI4K), and downstream effectors (Grb-2, PKCα, ERK, Akt, Src, Syk, ILK, ezrin) into confined segments of the cell membrane (‘docks’), that recursively assemble over hundreds of seconds. Average size (25±5 μm; ~80,000 individual analyses) and lifetime (222±25 sec) of docks extend by orders of magnitude previously recognized space-time frames for cell signaling. Activated/phosphorylated kinases clustered at membrane docks (99% P-Erb-B4; 91% P-Ret; 90% P-PKCα; 90% P-Src). STED super-resolution microscopy revealed up to 10 µm2 signal transducer areas of effector colocalization. Docks >1.0 µm2 accounted for ~90% of colocalizations versus ≈1% of raft-sized domains (<0.005 µm2). FRET-FLIM subsequently indicated that inter-molecular signal transducer interactions are essentially confined at docks. Docks were induced by growth factors (FGF-1, PDGF, EGF>HGF, IGF-1>>SCF/VEGF), were inhibited by Tyr kinase and cytoskeleton inhibitors, and were shown to induce cancer cell proliferation.

Citation Format: Saverio Alberti, Valeria Relli, Romina Tripaldi, Pasquale Simeone, Emanuela Guerra, Andrea Sacchetti, Paolo Ciufici, Antonino Moschella, Valeria Caiolfa, Moreno Zamai, Marco Trerotola. Growth factor-induced cell membrane macroplatforms drive protein kinase signaling for transformed cell growth [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 2550.
Abstract 2624: Assay toolkit for the discovery of new RAS pathway inhibitors
Authors: Kouznetsova Ekaterina , McCauley Lauren S. , Eells Rebecca , Steinke-Neal Elaine , Sullivan Timothy , Primavera Louis , Longen Charles , Ma Haiching
Published: April 2023

The small GTPase KRAS is a known oncogene that is very frequently mutated in a variety of cancers. Over the past several years, there has been a resurgence of interest in KRAS drug discovery following the identification of new inhibitor target sites and several molecules entering clinical trials.

We have produced proteins and set up assays for a large collection of Ras variants, guanine nucleotide exchange factors and effectors to facilitate drug discovery activities targeting KRAS/MAPK pathway. RBC’s extensive kinase assay portfolio enables identification of kinase inhibitors downstream of Ras. This poster will summarize our most recent assay development efforts for biochemical and biophysical assays for these targets.

Citation Format: Ekaterina Kouznetsova, Lauren S. McCauley, Rebecca Eells, Elaine Steinke-Neal, Timothy Sullivan, Louis Primavera, Charles Longen, Haiching Ma. Assay toolkit for the discovery of new RAS pathway inhibitors [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 2624.
Abstract 2644: BL-M02D1, a novel Trop2-targeting ADC, demonstrates robust anti-tumor efficacy in preclinical evaluation
Authors: Zhang Yong , Wan Weili , Zhuo Shi , Chen Lan , Li Gangrui , Zhao Shuwen , Khalili Jahan Salar , Xiao Sa , Yan Yongqi , Shen Xuejiao , Zhu Yi
Published: April 2023

Trop2, also known as trophoblast antigen 2, is a transmembrane glycoprotein. It is therapeutically targeted in cancer due to its over-expression in a variety of human carcinomas. To develop a promising therapeutic anti-tumor agent, we generated BL-M02D1, an anti-Trop2-Ed-04 ADC. It is comprised of a novel monoclonal antibody against Trop2 (hu4D3), a cathepsin B cleavable linker, and a novel topoisomerase I inhibitor agent (Ed-04). The novel Ed-04 is a derivative of the alkaloid camptothecin and mediates cell cycle arrest at the S phase and subsequent apoptosis. BL-M02D1 achieves a high drug-to-antibody-ratio (DAR=8) with a highly stable linker.

The antitumor efficacy of BL-M02D1 was evaluated in comparison to a commercialized Trop2-targeting ADC, IMMU-132, in xenograft tumor models. BL-M02D1 exhibited stronger tumor inhibition capacity than IMMU-132 at lower doses in the gastric cancer cell line NCI-N87, the breast cancer cell line MDA-MB-231, and the non-small cell lung cancer cell line HCC827 xenograft models. BL-M02D1 exhibited potent bystander effects, exemplified by strong tumor inhibition in a heterogeneous xenograft model of Trop2-positive and Trop2-negative tumor cells (A431 and SW620). This characteristic of BL-M02D1 was also compared to IMMU-132. In the heterogeneous Trop2 xenograft model (A431 and SW620), BL-M02D1 exhibited higher tumor inhibition capacity than IMMU-132, indicating that BL-M02D1 possess a more potent bystander effect than IMMU-132.

In summary, these studies suggest BL-M02D1, a novel Trop2-targeting ADC, is potentially more efficacious in the treatment of Trop2-expressing carcinomas than IMMU-132. The clinical phase I has been progressing and the available data exhibit excellent efficacy in breast cancer therapy with manageable toxicity.

Citation Format: Yong Zhang, Weili Wan, Shi Zhuo, Lan Chen, Gangrui Li, Shuwen Zhao, Jahan Salar Khalili, Sa Xiao, Yongqi Yan, Xuejiao Shen, Yi Zhu. BL-M02D1, a novel Trop2-targeting ADC, demonstrates robust anti-tumor efficacy in preclinical evaluation [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 2644.
Abstract 2681: Combination therapy of heme inhibitory protein (HeSP2) and Cyclopamine tartrate (CycT) with chemotherapeutic drugs is an effective strategy for treatment of TNBC
Authors: Alemi Parinaz Sadat , Castro Maria del Carmen Chacon , Berisha Eranda , Akter Zakia , Zhang Li
Published: April 2023

Breast cancer (BC) is the second leading cause of cancer death in women. Among the four main types of BC, triple-negative breast cancer (TNBC) accounts for 10 to 15% of BC. Although most TNBC patients have a poor prognosis, there are subsets of patients with tumors that respond well to chemotherapy. However, the outcome of chemotherapy in TNBC worsens over time due to the high expression of the oxidative phosphorylation (OXPHOS) complex. Likewise, drug resistant TNBC cells depend on OXPHOS; targeting oxidative metabolism and mitochondrial respiration can potentially overcome their drug resistance. Our lab recently engineered a heme-sequestering protein 2 (HeSP2) based on HasA hemophore. HeSP2 can effectively inhibit heme uptake and decrease the OXPHOS levels in cancer cells due to its high affinity for heme. Another agent, cyclopamine tartrate (CycT), previously known as a modulator of hedgehog signaling and mitochondrial respiration, can inhibit heme synthesis and reduce the OXPHOS levels. Therefore, using these heme-targeting agents in combination with chemotherapy could be an effective strategy to increase the therapeutic efficacy in TNBC. Cisplatin and etoposide are two chemotherapeutic drugs that have recently been used for treating TNBC. For this purpose, we examine the combination therapy of our OXPHOS inhibitory agents, HeSP2 and CycT, with cisplatin and etoposide in TNBC. In vitro experiment using three different TNBC cell lines, 4T1-Fluc-Neo, EMT6_Fluc_Puro, and MDA-MBA-231 confirmed the inhibitory effect of both HeSP2 and CycT in cell proliferation and colony formation in combination with or without chemo drugs. Results indicate that combination therapy significantly reduced the proliferation and inhibited colony formation in TNBC cells. As HeSP2 and CycT are therapeutic agents that reduce hypoxia and OXPHOS, they may also delay the emergence of drug resistance in cancer cells treated with chemotherapy. Our results indicate that HeSP2 and CycT in combination with chemotherapy drugs could serve as a promising therapy to diminish the tumorigenic function in TNBC and overcome resistance to chemotherapy.

Citation Format: Parinaz Sadat Alemi, Maria del Carmen Chacon Castro, Eranda Berisha, Zakia Akter, Li Zhang. Combination therapy of heme inhibitory protein (HeSP2) and Cyclopamine tartrate (CycT) with chemotherapeutic drugs is an effective strategy for treatment of TNBC [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 2681.
Abstract 2712: Killing cancer stem cells in 72 hours can cure breast cancer
Authors: Song Keum-soo , Nimse Satish Balasaheb , Kim Taisun
Published: April 2023

Advancements in stem cell technology can open new doors for treating various diseases, disorders, and disabilities, including cardiovascular diseases, gastrointestinal diseases, liver diseases, arthritis, neurological disorder, and baldness. Though stem cell technology offers several answers to treat these ailments, the success of this technology is unfortunately daunted by the development of stem cells into cancer stem cells (CSC) and eventually into cancer cells. CSCs are hypothesized to be key drivers of cancer cells and apparent drug resistance to the current anticancer drugs. Therefore, a drug that can kill the CSC within 72 hours can open a new window of success for stem cell therapy for treating severe diseases to a much simpler disorders such as baldness.

Herein, we present the application of the recently reported ID-Checker technology (Journal of Medicinal Chemistry 2022 65 (19), 12883-12894) for killing CSC. ID-Checker possesses a glucose tag that ensures its highly specific absorption and macroscale delivery of anticancer agents into the CSCs through GLUT channels. ID-Checker kills the cancer cells within 72 hours. ID-Checker inhibits the microtubule formation and GLUT channel deployment in 6 h and kills the CSCs without damaging normal cells.

Citation Format: Keum-soo Song, Satish Balasaheb Nimse, Taisun Kim. Killing cancer stem cells in 72 hours can cure breast cancer [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 2712.
Abstract 2760: Evaluation of functional heterogeneity in the potent responses of single CAR-T effector cells
Authors: Yellen Benjamin
Published: April 2023

Over the last decade, chimeric antigen receptor (CAR)-T cell immunotherapies have produced some miraculous cures in advanced stage, aggressive leukemias and lymphomas in both children and adults. However, many barriers continue to limit the availability and effectiveness of these therapies, including the lack of reliable potency assays for these cell-based “living” medicines. Unlike molecular medicines, which have well established manufacturing processes, cell based medicines are inherently heterogeneous, and each engineered lot of cells will inevitably have a distribution of growth properties, cytolytic activity, gene expression patterns, and secretion profiles, and it is not clear which cell subpopulations are most correlated with positive clinical outcomes.Here we report results of an evaluation of phenotypic heterogeneity in a population of anti-CD19 CAR-T cells co-cultured with target cells in an array of >300,000 microwells. Our approach uses high throughput time lapse imaging to continually monitor the cell-cell interactions between effector and target cells. We maintained cell viability in the microwells for more than 5 days, which gave us sufficient time to observe the full range of antigen-mediated cell killing phenotypes, and also evaluate growth rates of the potent effector cells. We tracked thousands of individual co-cultures through time lapse imaging, and cell behavior was evaluated and sorted by deep learning algorithms. We were able to quickly identify and retrieve clones that exhibited desirable phenotypes for full molecular characterization.Our analysis shows that three fundamental subpopulations of effector cells can be clearly distinguished and quantified. These are 1) effector cells that kill the targets and then die, 2) effector cells that kill the target and then proliferate, and 3) effector cells that do not kill the target. We further dissect these populations by characterizing their secretion and transcriptomic profiles, to identify further subtype classifications.

Citation Format: Benjamin Yellen. Evaluation of functional heterogeneity in the potent responses of single CAR-T effector cells [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 2760.
Abstract 278: Role of ATPIF1 in glioblastoma cell survival and metabolism
Authors: Guda Maheedhara Reddy , Mnatsakanyan Nelli , Morris Daniel , Asuthkar Swapna , Tsung Andrew J. , Velpula Kiran
Published: April 2023

The ATPase inhibitory factor 1 (ATPIF1) is a 12 kDa protein that inhibits the mitochondrial F1Fo-ATP synthase in the F1domain. It reverses the mitochondrial ATP synthase complex's function, contributing to the onset of oncogenesis. The overexpression of ATPIF1 in various cancers has recently attracted greater attention; however, its biological significance in human pathology, specifically cancer, remains unknown. Based on immunohistochemistry and Western blot analysis of glioblastoma (GBM) patient specimens, we found ATPIF1 is highly expressed in GBMs. Here we showed that mitochondrial fractions of silencing ATPIF1 in U-87 and U251 cells showed decreased expression of ATPIF1, ATP5A, and ATPG1-G2-G3. Next, we showed that manipulation of ATPIF1 showed increased Reactive Oxygen Species (ROS) and Caspase3/7 activity levels compared to untreated cells. ATPIF1 knockdown further caused cell cycle arrest at the G₀/G1stage, and FITC Annexin V analysis showed increased apoptosis. Increased glucose uptake is associated with aggressive oncological behavior, and here we found elevated levels of TCA metabolites in ATPIF1 knockdown cells. ATPIF1 silenced cells had a shift in extracellular acidification rate, accompanied by a simultaneous increase in proton leak, according to the metabolic analysis performed by Seahorse bio-analyzer. Overall, these observations suggest ATPIF1 plays a key role in the progression of GBM. However, further research is needed to verify the precise mechanisms by which ATPIF1 functions in GBM.

Citation Format: Maheedhara Reddy Guda, Nelli Mnatsakanyan, Daniel Morris, Swapna Asuthkar, Andrew J. Tsung, Kiran Velpula. Role of ATPIF1 in glioblastoma cell survival and metabolism [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 278.
Abstract 2782: Small molecule H182 suppresses Stat3 activation in tumor cells and combines with radiation therapy to block breast tumor growth in mouse syngeneic models
Authors: Chen Yue , Yue Peibin , Fu Wenzhen , Chen Weiliang , Kershaw Kathleen M. , Shiao Stephen L. , Tius Marcus A. , Lopez-Tapia Francisco , Turkson James
Published: April 2023

Signal transducer and activator of transcription 3 (Stat3) is a latent transcription factor that contributes to tumor cell growth and survival in constitutively-active form in several types of human cancers, and hence, serves as a therapeutic target. The azetidine-based compound, H182 irreversibly binds to Stat3. In cell-free DNA-binding assay, H182 selectively inhibited Stat3 DNA-binding activity (IC50 0.38-0.66 μM) over Stat1 or Stat5 (IC50>15.8 μM) in vitro. In treated pancreatic cancer cells, H182 specifically blocked the association of Stat3 with gp130 and JAK2, and inhibited Stat3 tyrosine phosphorylation and DNA-binding activity. Coimmunoprecipitation and colocalization studies of hemagglutinin (HA)-tagged Stat3 and EGFP-tagged Stat3 expressed in prostate cancer cells showed that treatment with H182 blocked the HA-Stat3:EGFP-Stat3 interactions in intact cells. Immunofluorescence staining with laser-scanning confocal microscopy analysis for the intracellular localization of Stat3 showed treatment of H182 disrupted Stat3 nuclear accumulation and promoted the aggregation of Stat3 at the perinuclear region. H182 consequently suppressed Stat3-dependent transcriptional activity and the expression of Stat3 downstream genes, including Cyclin A, Bcl-2, Cyclin B1, and Mcl-1. Moreover, H182 significantly inhibited the colony survival, migration, and invasion in vitro of breast, pancreatic and prostate cancer cells harboring aberrant Stat3 activation. Significantly, in vivo administration of H182 in combination with radiation induced a strong antitumor response against mouse triple-negative breast cancer in syngeneic models and prolonged survival. Thus, our study provides a novel Stat3 inhibitor with significant antitumor activity against human tumors cancer harboring persistently active STAT3.

Citation Format: Yue Chen, Peibin Yue, Wenzhen Fu, Weiliang Chen, Kathleen M. Kershaw, Stephen L. Shiao, Marcus A. Tius, Francisco Lopez-Tapia, James Turkson. Small molecule H182 suppresses Stat3 activation in tumor cells and combines with radiation therapy to block breast tumor growth in mouse syngeneic 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 2782.
Abstract 2783: TET2-STAT3-CXCL5 nexus promotes triglyceride transfer from neutrophils to cancer cells to fuel lung adeno-to-squamous transdifferentiation
Authors: Xue Yun , Chen Yuting , Sun Sijia , Ji Hongbin
Published: April 2023

Phenotypic plasticity is a rising cancer hallmark and lung adeno-to-squamous transdifferentiation (AST) triggered by LKB1 inactivation is significantly associated with drug resistance. Mechanistic insights into AST are urgently needed to identify therapeutic vulnerability in LKB1-deficient lung cancer. Here we find that Ten-Eleven Translocation (TET)-mediated active DNA demethylation is elevated during AST in KrasLSL-G12D/+;Lkb1L/L(KL) mice, and knockout of individual Tet gene reveals that Tet2 is required for squamous transdifferentiation. STAT3 recruits TET2 to CXCL5 promoter region for demethylation and promotes neutrophil infiltration. Targeting the STAT3-CXCL5 nexus effectively inhibits squamous transdifferentiation through reducing neutrophil infiltration. Interestingly, tumor-infiltrating neutrophil is laden with triglycerides and transfer lipid to tumor cells to promote cell proliferation and squamous transdifferentiation. Pharmacological inhibition of macropinocytosis dramatically inhibits neutrophil-to-cancer cell lipid transfer and blocks squamous transdifferentiation. These data uncover an epigenetic mechanism orchestrating phenotypic plasticity through regulating immune microenvironment and metabolic communication, and identify therapeutic strategies to inhibit AST.

Citation Format: Yun Xue, Yuting Chen, Sijia Sun, Hongbin Ji. TET2-STAT3-CXCL5 nexus promotes triglyceride transfer from neutrophils to cancer cells to fuel lung adeno-to-squamous transdifferentiation [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 2783.
Abstract 2826: Electromagnetic fields and hyperthermia induce lysosomal permeabilization and death in different cancer cell models: in vitro and in vivo studies
Authors: Obrador Elena , Jihad-Jebbar Ali , Salvador-Palmer Rosario , López-Blanch Rafael , Oriol-Caballo María , Moreno-Murciano María Paz , Navarro Enrique , Cibrián Rosa , Estrela Jose M.
Published: April 2023

Introduction: Non-ionizing radiations (NIR), e.g. tumor-treating fields, high intensity focused ultrasounds (HIFU) or high frequency microwaves, have potential applications in Oncotherapy. Nevertheless, as today, their efficacy is limited. We investigated whether externally applied magnetic fields and non-ablative hyperthermia affect cancer cell viability and growth.

Methods: Under in vitro conditions, human A2058 melanoma, AsPC1 pancreas carcinoma, and MDA-MB-231 breast carcinoma cells were treated with electromagnetic fields (EMFs, using solenoids) and/or hyperthermia (HT, using a thermostated water bath). The effect of both energies was also studied in combination with standard chemotherapy and targeted therapy. An experimental system combining EMFs and HIFU-induced HT was developed and tested in vivo.

Results: EMFs (25 μT, 4h) or HT (52ºC, 40 min) quite moderately affect cancer cell viability. However, their combination decreased viability to approx. 16%, 50%, and 21% of controls values in A2058, AsPC1, and MDA-MB-231 cells, respectively). Increased lysosomal permeability, release of cathepsins into the cytosol and activation of the mitochondria-dependent cell death are the underlying mechanisms. Cancer cells were completely eliminated combining EMFs, HT and standard chemotherapy or EMFs, HT and anti-Hsp70-targeted therapy. In vivo experiments, using pancreatic cancer xenografts growing in nude mice, show that combination of EMFs, HIFU-induced HT and gemcitabine induces a drastic cancer regression (to approx. 3-4% of control values).

Conclusion: Non-invasive NIR (EMFs and HIFU-induced HT) can complement current Oncotherapies. This strategy, associated to current chemotherapy or anti-Hsp70-targeted therapy, may facilitate the elimination of tumors that can be previously located using standard imaging techniques (CT scan and NMR). These results may help to overcome the limitations in the use of different NIR in Oncotherapy.

Citation Format: Elena Obrador, Ali Jihad-Jebbar, Rosario Salvador-Palmer, Rafael López-Blanch, María Oriol-Caballo, María Paz Moreno-Murciano, Enrique Navarro, Rosa Cibrián, Jose M. Estrela. Electromagnetic fields and hyperthermia induce lysosomal permeabilization and death in different cancer cell models: in vitro and in vivo studies [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 2826.
Abstract 2827: Application of AZD2281 combine with radiation therapy in glioblastoma
Authors: Liao Tsal-Lan , Chiang I-Tsang , Chang Yuan , Hsu Fei-Ting
Published: April 2023

Glioblastoma (GBM), can be divided into primary tumors from brain tissue (>90% of all cases) and secondary tumors that progress from low-grade gliomas. The current GBM treatment strategy is chemotherapy drug temozolomide (TMZ) combined with radiotherapy (RT), which has limited treatment effect, poor prognosis, high recurrence rate, and short prognosis survival time. Recently, studies indicated that breast cancer susceptibility gene 1/2 (BRCA1/2) mutations are frequently occur in GBM, while few studies have explored this pathway in depth. At present, AZD2281 (PARP Inhibitor) has been used to treat breast cancer and ovarian cancer patients with BRCA1/2 mutation...etc. Whether AZD2281 can play a role in affecting the relevant pathways after BRCA1/2 mutate GBM patients, and the possible mechanism and benefit of combined RT are the focus of this study. AZD2281 is the first Poly (ADP-ribose) polymerase (PARP) inhibitor approved by the U.S. Food and Drug Administration (FDA) for the treatment of germline BRCA mutation-associated ovarian cancer. AZD2281 is a targeted therapy that used to block DNA Damage Response (DDR) in cells/tumors deficient in homologous recombination repair (HRR). After RT, there may be a phenomenon of DNA repair; therefore, AZD2281 can be used to disrupt the repair process. At the beginning, we used RSI analysis to confirm that patient with higher expression level of BRCA1 may relatively resistance to RT. To overcome this issue, we identified the effect of AZD2281 combined with RT on GBM cells (BRCA1/2 higher and lower expression cells) by (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay), and colony formation. The internal/external apoptosis pathways (Caspase-3/Caspase-8/Caspase-9) and TUNEL assay were used to prove the superior apoptosis effect of AZD2281 combined with RT as compared to single treatment. Additionally, cells lower BRCA1 expression level (U87 and U251 siBRCA1) may sensitive to RT treatment. In our DNA damage, DNA fragmentation, and transwell invasion/migration results, we showed that the combination of AZD2281 and RT may effective induce DNA damage. In our animal data, we proved that combination therapy can effective suppression tumor growth and apoptosis-related protein signaling increased in DNA damage by IHC (p-ATM/p-CHK2/Caspase-3/Caspase-8/Caspase-9). In conclude, we suggested that AZD2281 can be used as a radiosensitizer to sensitize GBM patient to RT, which is dependent to BRCA1 expression.

Citation Format: Tsal-Lan Liao, I-Tsang Chiang, Yuan Chang, Fei-Ting Hsu. Application of AZD2281 combine with radiation therapy in glioblastoma [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 2827.
Abstract 2836: H2S donors as radiosensitizers for glioblastoma multiforme (GBM)
Authors: Modi Morni , Weber Camila , Xu Qinqin , Chen Kuanling , Cheong Ana , Nagel Zachery , Harrison Lynn
Published: April 2023

Glioblastoma multiforme (GBM) is an extremely aggressive CNS cancer in adults. Radiation is a cornerstone therapy but poses high risk of healthy tissue necrosis. Despite maximal treatment, GBM results in death within 15-21 months of diagnosis. These poor statistics suggest the need for a novel therapy. The use of radiosensitizing agents is a novel approach to minimizing radiation mediated healthy tissue necrosis by allowing use of lower doses to achieve comparable cytotoxic effects. Our lab has shown that sodium sulfide (Na2S), a hydrogen sulfide (H2S) donor, selectively kills GBM (T98G) cells while sparing normal human brain endothelial (hCMEC/D3) cells. Na2S also selectively radiosensitizes GBM cells and enhances cell killing with photon or proton radiation. We are currently exploring Diallyl Trisulfide (DATS), another H2S donor, as a radiosensitizing agent and have demonstrated that DATS can kill GBM cells. At a given dose of 2 Gy, radiation alone decreases T98G survival by 10%. With DATS, survival decreases by 70%, suggesting an additive effect. To determine if DATS increases DNA damage mediated cell death, T98G cells were treated with DATS, radiation, or combined DATS + radiation treatment and then analyzed for γH2AX foci, an indicator of double stand breaks (DSB). Our results show that compared to no treatment, DATS increased the number of γH2AX foci per cell by 1.5-fold, by 3-fold with radiation, and by 4-fold with combined DATS + radiation, an additive response. To elucidate the mechanism of increased DNA damage, a novel fluorescence multiplex host cell reactivation (FM-HCR) reporter system was used to assess repair activity of DSBs in live cells. T98G cells pre-treated with DATS were transfected with reporter plasmids containing site-specific lesions, mimicking radiation-like DNA damage. Repair resulted in expression of a fluorescent reporter protein, which was quantitated using flow cytometry. Our results show that DATS did not alter non-homologous end joining (NHEJ) or homologous repair (HR) activity of DSB. However, DATS did alter base excision repair (BER) activity by decreasing repair of abasic lesions and increasing repair 8oxoG lesions. We hypothesize that this attempted BER via simultaneous strand cleavage by AP endonuclease (Ape1) or glycosylase (Ogg1) may lead to build up of repair intermediates, threatening genomic stability and inducing DSB. Pursuing specific DNA repair biochemical assays is required to further understand this effect of DATS on DNA damage and repair inhibition.

Citation Format: Morni Modi, Camila Weber, Qinqin Xu, Kuanling Chen, Ana Cheong, Zachery Nagel, Lynn Harrison. H2S donors as radiosensitizers for glioblastoma multiforme (GBM) [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 2836.
Abstract 2838: Tumor-targeted interleukin-2 enhances the anti-cancer activity of multivalent OncoFAP radioligand therapeutics
Authors: Galbiati Andrea , Bocci Matilde , Gilardoni Ettore , Zana Aureliano , Neri Dario , Cazzamalli Samuele
Published: April 2023

Radioligand Therapeutics (RLTs) are products composed of a tumor-targeting small organic ligand and a biocidal radioactive payload. The therapeutic potential of RLTs strongly depends on their uptake and residence time in tumors and in the healthy structures after administration. Fibroblast Activation Protein (FAP) is a cell surface antigen abundantly expressed in the microenvironment of the majority of solid human cancers of epithelial origin. We have recently described OncoFAP, an ultra-high affinity ligand of FAP whose targeting performances have been validated by Nuclear Medicine in patients with a wide variety of solid malignancies. Multivalent OncoFAP radioligand therapeutics (OncoFAP-RLTs) show high and prolonged tumor uptake with favorable tumor-to-organ ratios. Here we describe the development of novel multivalent OncoFAP-RLTs and their combination with a clinical-stage tumor-targeted interleukin-2 (L19-IL2). We designed and synthesized a set of multivalent OncoFAP derivatives, aiming at maximizing their affinity for the target protein and their residence time in the catalytic pocket of FAP. The multivalent ligands were coupled to the DOTAGA chelator, suitable for radiolabeling with 177Lu for therapeutic application. The tumor targeting properties of the multivalent OncoFAP ligands were investigated in a comparative biodistribution study in SK-RC-52.hFAP tumor-bearing mice. 177Lu-OncoFAP-23 emerged for its superior uptake and residence time in tumors and for its favorable tumor-to-kidney ratio. Moreover, when administered at very low dose 177Lu-OncoFAP-23 exhibited an enhanced anticancer activity compared to a monovalent (177Lu-OncoFAP-DOTAGA) and bivalent (177Lu-BiOncoFAP-DOTAGA) version of OncoFAP. When OncoFAP-based RLTs were combined with L19-IL2, a strong synergistic anticancer activity was observed. This synergy is driven by NK-cell mediated immune response directed to the tumor, with the concomitant release of granzymes and other pro-apoptotic proteins in the tumor microenvironment. Our findings heighten OncoFAP-based RLTs as promising anti-cancer drugs for treatment of FAP-positive tumors, alone or in combination with targeted interleukin-2.

Citation Format: Andrea Galbiati, Matilde Bocci, Ettore Gilardoni, Aureliano Zana, Dario Neri, Samuele Cazzamalli. Tumor-targeted interleukin-2 enhances the anti-cancer activity of multivalent OncoFAP radioligand therapeutics [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 2838.
Abstract 299: Large tandem duplications in cancer resulting from transcription and DNA replication collisions
Authors: Yang Yang , Badura Michelle , O’Leary Patrick , Egusa Emily , Robinson Troy , Zhong Xiaoming , Swinderman Jason , Kim Minkyu , Li Haolong , Ashworth Alan , Feng Felix , Chou Jonathan , Yang Lixing
Published: April 2023

Somatic structural variations (SVs) are common in cancer. Although a small fraction of SVs in breast and ovarian cancers can be attributed to homologous recombination deficiency, the underlying molecular mechanisms for the vast majority of somatic SVs remain unclear. Here, we focus on the roles of transcription and DNA replication collisions in genomic instability in cancer. Such collisions are unavoidable in cells since both transcription and replication use the same DNA as template. We hypothesized that transcription replication collisions (TRCs), if not properly repaired, would lead to collapsed replication forks and result in SVs. To this end, we studied somatic SVs in 5994 high-coverage whole-genome sequenced primary and metastatic tumors from three independent pan-cancer cohorts. A total of 12 conserved SV signatures, representing independent molecular mechanisms, were deconvoluted from these cohorts using non-negative matrix factorization approach. We detected replicated-strand bias, the expected footprint of transcription-replication collision, in large tandem duplications (TDs) across multiple cohorts. This bias was only observed in expressed genes, consistent with TRCs depending on transcription activity. Large TDs were abundant in female-specific (breast, ovarian and uterus), upper gastric-intestinal tract and prostate cancers. They were associated with worse patient survival and TP53 and CDK12 mutations. CDK12 is a cyclin-dependent kinase (CDK) and a key regulator of transcription elongation. Deleting or suppressing CDK12 using CRISPR-Cas9 in prostate cancer cell lines not only increased RNA:DNA hybrids (R-loops), but also promoted TRCs, suggesting a mechanism by which dysregulation of a transcriptional CDK may lead to genomic instability. Finally, using existing large-scale drug screening data, we found that cancer cell lines with abundant large TDs were significantly more sensitive to the WEE1 inhibitor, MK-1775, which we experimentally validated in prostate cancer cells lacking CDK12. In summary, our data suggest that large TDs in cancer form due to impaired TRC repair and can be used as a biomarker for prognosis and treatment.

Citation Format: Yang Yang, Michelle Badura, Patrick O’Leary, Emily Egusa, Troy Robinson, Xiaoming Zhong, Jason Swinderman, Minkyu Kim, Haolong Li, Alan Ashworth, Felix Feng, Jonathan Chou, Lixing Yang. Large tandem duplications in cancer resulting from transcription and DNA replication collisions [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 299.
Abstract 2993: Unmutated tumor antigens are abundant and contribute to tumor control in melanoma
Authors: Apavaloaei Anca , Zhao Qingchuan , Hesnard Leslie , Vincent Krystel , Hardy Marie-Pierre , Durette Chantal , Lanoix Joël , Laverdure Jean-Philippe , Larouche Jean-David , Cuevas Maria Virginia Ruiz , Ehx Grégory , Lemieux Sébastien , Thibault Pierre , Perreault Claude
Published: April 2023

Recognition of MHC-I-associated tumor antigens (TAs) by CD8+ T cells is central to antitumor immunity. Owing to the elevated tumor mutational burden (TMB) in melanoma, the marked efficacy of immune checkpoint blockade (ICB) has been attributed to the recognition of mutated TAs. However, recent reports showed that response to ICB in melanomas with low TMB is associated with CD8+ T-cell reactivity against melanocyte lineage-associated antigens (LSAs). Here, we systematically evaluated the contribution of all TA classes, i.e., mutated and unmutated, canonical and non-canonical, to the antigenic landscape of melanoma. We characterized the TAs from melanoma biopsies and patient-derived cell lines using proteogenomics. Out of 79450 MHC-I-associated peptides (MAPs) identified from 19 samples, we found 557 unmutated TAs classified as tumor-specific (TSA), tumor-associated (TAA), or LSAs. These TAs most often derived from annotated open-reading frames, followed by ncRNAs and intergenic regions. By contrast, only 6 MAPs were mutated and tumor-specific, which could be partially explained by a decreased expression of mutations within MAP-generating genomic regions. While the number of unmutated TAs with predicted presentation (TApres) in melanoma patients was similar between responders and non-responders pre-ICB, non-responders showed marks of inefficient antigen presentation. In consequence, only responders lost TApres upon treatment, in tandem with an expansion in tumor-infiltrating lymphocytes. These results reveal a previously underappreciated contribution of unmutated TAs to tumor control in melanoma and suggest that enhancing their recognition could improve the ICB efficacy in non-responders.

Citation Format: Anca Apavaloaei, Qingchuan Zhao, Leslie Hesnard, Krystel Vincent, Marie-Pierre Hardy, Chantal Durette, Joël Lanoix, Jean-Philippe Laverdure, Jean-David Larouche, Maria Virginia Ruiz Cuevas, Grégory Ehx, Sébastien Lemieux, Pierre Thibault, Claude Perreault. Unmutated tumor antigens are abundant and contribute to tumor control in melanoma [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 2993.
Abstract 2996: CT26 neoantigen presentation and immunogenicity
Authors: Nyayapathy Seeta , Jones Richard , Ford Michael , Doty Micah , Rumble Julie M.
Published: April 2023

The foundation of personalized cancer therapy is teaching a patient’s own immune system to directly kill the cancer cells. Vaccination against tumor antigens is one method being explored to generate immune responses against tumors, and may be particularly effective in conjunction with therapies currently in the clinic, such as checkpoint inhibition. Identification of antigens for use in a vaccine is the pinch point of this method, and many algorithms have been developed in an attempt to predict what epitopes will be both presented and immunogenic. Importantly, both public (common to multiple individuals/cancer types) and private (single individual) epitopes may contribute to the development of effective vaccines. Experimental identification of presented and immunogenic epitopes is crucial to building effective anti-cancer vaccines. Using immunopeptidome profiling, we have narrowed a collection of published potential neoantigens to those which were presented by MHC class I in the CT26 mouse colorectal cancer cell line. These peptides were synthesized and used to immunize BALB/c mice. The immunogenicity of the peptide mix was then evaluated by ELIspot to assess IFN-gamma responses in immunized mice. These experiments support a complete workflow for selection of potential cancer neoantigens which experimentally exhibit both MHC presentation and immunogenicity.

Citation Format: Seeta Nyayapathy, Richard Jones, Michael Ford, Micah Doty, Julie M. Rumble. CT26 neoantigen presentation and immunogenicity [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 2996.
Abstract 3012: Increased lung cancer risk and associated risk factors in tuberculosis survivors: a Korean population-based study
Authors: Park Dong Won , Moon Seong Mi , Choi Hayoung , Kim Sang Hyuk , Kang Hyung Koo , Jung Jin Hyung , Han Kyungdo , Shin Dong Wook , Lee Hyun
Published: April 2023

Background: Although studies have suggested an association between previous tuberculosis (TB) infection and increased lung cancer risk, there are limited data comprehensively evaluating the effects of smoking and chronic obstructive pulmonary disease (COPD) on the association.

Methods: This population-based cohort study enrolled TB survivors (n=75,467) ≥20 years of age between 2010-2017 and 1:1 age- and sex-matched controls (n=75,467). Subjects were followed for one year after the date of TB diagnosis to the date of the incidental lung cancer, death, or December 2018.

Results: During a median follow-up duration of 4.8 (interquartile range, 2.9-6.6) years, the incident lung cancer risk was 1.71-fold (95% confidence interval [CI], 1.49-1.97) higher in TB survivors than in the controls. Compared to never-smoker, non-TB infected controls, TB survivors who were current smokers with ≥20 pack-years showed the highest risk of lung cancer (adjusted hazard ratio [aHR], 6.78; 95% CI, 5.26-8.74). In terms of COPD, the risk of lung cancer was higher in TB survivors with COPD compared to non-COPD, non-TB-infected controls (aHR, 2.43; 95% CI, 2.03-2.91). However, smoking as well as COPD did not have significant interaction on the association between TB and lung cancer development (p for interaction>0.05 for both). Risk factors related to incident lung cancer among TB survivors were pulmonary TB, age >60 years, smoking, and the presence of COPD or asthma.

Conclusions: Previous TB infection is an independent risk factor for lung cancer without significant interaction with smoking status or amount and COPD.

Citation Format: Dong Won Park, Seong Mi Moon, Hayoung Choi, Sang Hyuk Kim, Hyung Koo Kang, Jin Hyung Jung, Kyungdo Han, Dong Wook Shin, Hyun Lee. Increased lung cancer risk and associated risk factors in tuberculosis survivors: a Korean population-based study [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 3012.
Abstract 3031: Pilot study of EGFR inhibition with erlotinib in liver fibrosis for hepatocellular carcinoma prevention
Authors: Tanabe Kenneth K. , Zahrieh David , Strand Carrie A. , Hoshida Yujin , Flotte Thomas J. , Della’Zanna Gary , Umar Asad , Limburg Paul
Published: April 2023

BACKGROUND: Effective approaches for prevention of hepatocellular carcinoma (HCC) will have a significant impact on HCC-related mortality. Data from preclinical models strongly support epidermal growth factor receptor (EGFR) as a target for chemoprevention of HCC. Erlotinib is a small-molecular EGFR tyrosine kinase inhibitor that is FDA-approved for cancer treatment. The side effects of erlotinib observed in patients receiving the oncology dose (150 mg/day) render it unacceptable as a chemoprevention agent at this dose.

OBJECTIVE: The objective of this clinical trial was determination of a safe and minimum effective dose of erlortinib for which a ≥50% reduction phospho-EGFR immunohistochemical staining in the liver was observed.

RESULTS: 46 participants were pre-registered and 25 participants were registered in this multicenter trial. In a dose de-escalation trial design, cohorts of participants received a 7 day course of erlotinib 75 mg/day, 50 mg/day or 25 mg/day with liver tissue acquisition prior to and after erlotinib. A ≥50% reduction phospho-EGFR immunohistochemical staining in the liver was observed in a minimum of 40% of participants at each of the dose levels. Erlotinib was very well tolerated with few side effects observed, particularly at the dose of 25 mg/day. A favorable modulation of the Prognostic Liver Signature was observed in participants that received the higher erlotinib doses (50 and 75 mg/day).

CONCLUSION: These data support the selection a dose of 25 mg/day of erlotinib for a longer intervention to assess for evidence of efficacy as an HCC chemoprevention drug.

Citation Format: Kenneth K. Tanabe, David Zahrieh, Carrie A. Strand, Yujin Hoshida, Thomas J. Flotte, Gary Della’Zanna, Asad Umar, Paul Limburg. Pilot study of EGFR inhibition with erlotinib in liver fibrosis for hepatocellular carcinoma prevention [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 3031.