bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A032: Cell membrane coated- Biomimetic biodegradable nanoparticles for tumor targeted delivery of THP-doxorubicin using polylactic acid based redox responsive polymer
Authors: Kaur Harshdeep , Singh Harpal , Singh Neetu
Published: December 2023

Chemotherapy is a widely used systemic treatment modality for cancer treatment as it kills the rapidly dividing cells. However, the administered chemo-drugs cause systemic toxicity,  have fast degradation rate at physiological conditions hence low bioavailability and poor tumor accumulation. These shortcomings can be mitigated by the use of tumor targeting drug delivery systems. The nanoparticles can further be tailor made according to physiological conditions of the tumor microenvironment. Herein, the redox responsive polymer has been synthesized by introducing the disulphide linkage in the polymer. The drug release profile indicated the release of the drug is more efficient in presence of 10mM Dithiothreitol(reducing agent for disulphide linkage) when compared to 0mM dithiothreitol. In this context, this study investigates the therapeutic efficacy of THP-doxorubicin encapsulated in polylactic acid based redox responsive nanoparticle(NP) whose surface have been modified with tumor (4T1) and macrophage (J774) cell membranes. The coated cell membrane provides advantages such as immune escape, long blood circulation time, specific molecular recognition and cell targeting. The prepared nanoparticles have a hydrodynamic size  of 210±10 nm with zeta potential of -30±3 mV. The cellular internalization studies of free drug vs the cell membrane coated NPs (CMNPs) were done and it was confirmed that CMNPs were internalized more efficiently in one hour drug incubation study. The same was confirmed when cancer cell proliferation inhibition assay was done, and half maximal inhibitory concentration of free drug, uncoated NPs and CMNPs were 0.31µM , 0.463µM and 0.137µM respectively. This study provides the synthesis and characterization of redox responsive polymer, preparation of cell membrane coated nanoparticles and in-vitro studies for the same. This study has a future scope in the field of personalized medicine at much cheaper price when compared to active targeting methods by antibodies, CART, proteins etc.

Citation Format: Harshdeep Kaur, Harpal Singh, Neetu Singh. Cell membrane coated- Biomimetic biodegradable nanoparticles for tumor targeted delivery of THP-doxorubicin using polylactic acid based redox responsive polymer [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A032.
Abstract A042: Targeting vulnerabilities arising from global DNA hypomethylation in cancer
Authors: Mustafi Pallabi , Hanratty Brian , Coleman Ilsa , Patel Radhika , Mohamed Adil , Sarthy Jay , Ha Gavin , Haffner Michael
Published: December 2023

Background: DNA methylation alterations are a universal feature of cancer. In addition to site specific gain of DNA methylation (hypermethylation), a global loss of methylation (hypomethylation) was noted in most cancer genomes. Whereas numerous studies have focused on the role of hypermethylation in disease progression, less is known about the biology of hypomethylation in cancer. Recently, we have identified tumors across all major cancer types that are characterized by severe loss of DNA methylation. Our preliminary data suggest that these DNA hypomethylated cancers (hereafter DHMCs) show distinct alterations influencing gene expression and tumor microenvironment composition.

Objective: We hypothesize that since DNA methylation is critical for genome organization and gene expression. Severe global hypomethylation is likely to result in distinct epigenomic and genomic alterations. We therefore investigate common molecular changes associated with DNA hypomethylation to develop novel therapeutic approaches targeting unique vulnerabilities arising in DHMCs.

Study design: To delineate the biology of DHMCs, we determined methylation changes with validated orthogonal methods in large representative cohorts of patient samples (The Cancer Genome Atlas (TCGA), University of Washington rapid autopsy cohorts), patient derived xenografts and cancer cell lines. We assessed the patterns of common molecular alterations by performing genome wide epigenome mapping experiments and by defining common driver gene events in DHMCs. We further determined tumor cell intrinsic therapeutic vulnerabilities in broad pharmacologic and CRISPR-Cas9 genomic screens.

Results: We observed that DNA hypomethylation is a relatively common feature of solid tumors affecting overall 15-20% of all cancers, with highest rates observed in urothelial, melanoma, hepatocellular and lung and head and neck squamous cell carcinoma. In in vivo tumor models of DHMCs, we observe distinct shifts in epigenetic states upon DNA hypomethylation. Most notably, we found that DHMCs are characterized by unique intrinsic drug sensitivities to drug classes affecting core histone levels.

Impact: This is the first study to show druggable vulnerabilities arising from DNA hypomethylation. Collectively, these studies define a novel epigenetic subtype of cancer, determine its clinical and molecular features, and pave the way for new targeted therapies.

Citation Format: Pallabi Mustafi, Brian Hanratty, Ilsa Coleman, Radhika Patel, Adil Mohamed, Jay Sarthy, Gavin Ha, Michael Haffner. Targeting vulnerabilities arising from global DNA hypomethylation in cancer [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A042.
Abstract A053: The dual BRG1/BRM (SMARCA4/2) inhibitor FHD-286 induces functional differentiation and splicing defects in preclinical models of acute myeloid leukemia (AML)
Authors: Gartin Ashley K. , Mayhew David N. , Lahr David L. , Elliott GiNell , Gandevia Heena , Wilson Molly M. , Centore Richard , Collins Mike , Thomsen Astrid , Piel Jessica , Sandoval Gabriel J. , Hentemann Martin
Published: December 2023

The BRG/BRM associated factor (BAF) complex is a chromatin remodeler critical for maintenance of cell viability in many cancers. This includes acute myeloid leukemia (AML), where BAF maintains the stem-like transcriptional state of blast cells. FHD-286 is a dual inhibitor of the BAF ATPase subunits BRG1 and BRM and is currently being investigated in relapsed or refractory AML and myelodysplastic syndrome. We have shown previously that FHD-286 induces the expression of the myeloid differentiation marker CD11b in preclinical models after treatment with sub-cytoreductive doses at long timepoints. Here we further characterized the differentiation phenotype induced by low-dose FHD-286 in AML cell lines through RNA-seq and a genome-wide CRISPR-Cas9 knockout screen. These datasets and mechanistic follow-up studies suggested AML cell lines treated with FHD-286 can functionally differentiate to gain the ability to produce superoxide anion and perform phagocytosis. We also found that FHD-286 treatment disrupts mRNA splicing and that this is a likely contributor to AML cell growth defects induced by BRG1/BRM inhibition. This study suggests multiple mechanisms by which FHD-286 is able to disrupt the stem-like transcriptional state of AML blasts to cause differentiation and ultimately cell death.

Citation Format: Ashley K. Gartin, David N. Mayhew, David L. Lahr, GiNell Elliott, Heena Gandevia, Molly M. Wilson, Richard Centore, Mike Collins, Astrid Thomsen, Jessica Piel, Gabriel J. Sandoval, Martin Hentemann. The dual BRG1/BRM (SMARCA4/2) inhibitor FHD-286 induces functional differentiation and splicing defects in preclinical models of acute myeloid leukemia (AML) [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A053.
Abstract A055: Discovery of a novel, highly potent VHL-recruiting EZH2 PROTAC degrader targeting MLL-r AML
Authors: Velez Julia , Yu Xufen , Dale Brandon , Park Kwang-Su , Ümit Kaniskan H. , Jin Jian
Published: December 2023

Mutated or overexpressed Enhancer of Zeste Homolog 2 (EZH2), the catalytic subunit of the Polycomb Repressive Complex 2 (PRC2), has been implicated in numerous cancers, including breast, leukemia, and lung. Through its methyltransferase activity, EZH2 tri-methylates H3K27 to repress transcription. In addition to its enzymatic activity, recent studies have highlighted non-enzymatic oncogenic functions of EZH2 that limit the efficacy of enzymatic inhibitors. Therefore, alternative therapeutic strategies such as proteolysis targeting chimera (PROTAC) degraders that suppress both enzymatic and non-enzymatic activity have been developed. PROTACs are heterobifunctional molecules consisting of a protein of interest (POI) binder linked to an E3-ligase ligand. The manufactured proximity between the POI and the recruited E3-ligase complex initiates ubiquitination of the POI, resulting in its degradation via the 26S proteasome. EZH2 exhibits enzymatic and non-enzymatic oncogenic activity in various cancers including MLL1-rearranged acute myeloid leukemia (MLL-r AML) and triple negative breast cancer (TNBC). Here we present MS8847, a novel VHL-recruiting EZH2 PROTAC degrader. In MLL-r AML cells, MS8847 induces potent EZH2 degradation in a ubiquitin proteasome-, time- and concentration-dependent manner. Compared to the previously published EZH2-targeting PROTAC degraders (MS8815, YM281, U3i, and E7) and parent inhibitor, Tazemetostat, MS8847 displays superior EZH2 degradation and cell growth inhibition in MLL-r AML cell lines dependent on both enzymatic and non-enzymatic EZH2 activity. MS8847 also induces EZH2 degradation and cell growth inhibition in TNBC cell lines. Moreover, MS8847 has a favorable PK profile making it suitable for in vivo studies. Together, MS8847 offers an improved VHL-recruiting EZH2 PROTAC degrader with potential as an effective therapeutic agent to treat EZH2-dependent cancers.

Citation Format: Julia Velez, Xufen Yu, Brandon Dale, Kwang-Su Park, H. Ümit Kaniskan, Jian Jin. Discovery of a novel, highly potent VHL-recruiting EZH2 PROTAC degrader targeting MLL-r AML [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A055.
Abstract A059: Small molecule microarray lysate screen identifies bromodomain ligands that target the MYC transcription regulatory network
Authors: Cohen Emily B , Ma Zhihua , Glore Wayne L , Volkov Oleg A , Liang Tong , Obholzer Nikolaus D , Clay Andrew C , Pingili Mulini Reddy , Zheng Yupeng , Li Jun , Wang Shi Yun , Chen Tom , Freeman David B , Gao Hua , Zhou Minyun , Cheung Priscilla , Hopkins Tamara D , Pop Marius S , Dinsmore Christopher J , Trotter B. Wesley , Lin Charles Y , Rahl Peter B
Published: December 2023

MYC is the most frequently amplified gene in human cancers and one of the most sought-after drug targets for cancer therapy. Its function as a transcription factor is essential for its oncogenic potential. However, development of small molecules that target oncogenic MYC function in cells has been intractable due to its lack of known ligand binding sites. To overcome these structural challenges, we leveraged Kronos Bio’s small molecule microarray (SMM) screening platform to identify small molecules that can bind to MYC transcriptional complexes in cell lysates from cancer cells with deregulated MYC function. Unbiased transcriptional signature-based profiling identified multiple SMM hits that modulated the MYC transcription regulatory network (TRN) in MYC-dependent cancer cell lines. One of the SMM hits mimicked signature changes due to MYC loss of function, which were also similar to signature changes effected by BET bromodomain inhibitors. Biochemical, biophysical, and structural analyses found that the SMM hit could directly engage with BET bromodomains in purified systems. Cell-based proximity labeling demonstrated binding to BET proteins in live cells, and comparative pharmacology approaches suggested that the BET protein engagement was likely driving the observed gene expression changes. These results demonstrate the utility of the SMM platform in identifying ligands that modulate transcription factor TRNs through binding to critical cofactors.

Citation Format: Emily B Cohen, Zhihua Ma, Wayne L Glore, Oleg A Volkov, Tong Liang, Nikolaus D Obholzer, Andrew C Clay, Mulini Reddy Pingili, Yupeng Zheng, Jun Li, Shi Yun Wang, Tom Chen, David B Freeman, Hua Gao, Minyun Zhou, Priscilla Cheung, Tamara D Hopkins, Marius S Pop, Christopher J Dinsmore, B. Wesley Trotter, Charles Y Lin, Peter B Rahl. Small molecule microarray lysate screen identifies bromodomain ligands that target the MYC transcription regulatory network [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A059.
Abstract A079: FHD286 is a BRG1/BRM ATPase inhibitor showing efficacy in NSCLC models and is applicable to NSCLC patients both as a single agent treatment and in combination with current standards of care
Authors: Mikse Oliver , Adam Ammar , Elliott GiNell , Lahr Dave , Amaral Victoria , Viswanathan Priyadharshini , Johannessen Liv , Ahmad Hafiz , Wan Jessica , Bellon Steven , Hentemann Murphy
Published: December 2023

The BAF (BRG/BRM-associated factors) complex plays a critical role in shaping the chromatin landscape. The two mutually exclusive ATPases BRM/BRG1 (SMARCA2/4, respectively) drive chromatin remodeling activity in the BAF complex. Genetic alterations in the SMARCA4 gene, encoding BRG1, are found in approximately 10% of non-small cell lung cancer (NSCLC) patients, leaving this subset of BRG1-deficient tumors highly susceptible to inhibition of BRM (Hoffman, 2014; Oike, 2013). However, BRG1 can also have oncogenic activity in NSCLC (Li, 2019) and can serve as a predictive biomarker for chemotherapy outcome in patients (Bell, et al 2015), rationalizing the use of a dual BRG1/BRM ATPase inhibitor (FHD286) in NSCLC patients in combination with chemotherapy. Here, we demonstrate that a panel of in vitro and in vivo NSCLC models respond to BAF ATPase inhibition as a single agent treatment, irrespective of SMARCA4 mutation status. Analysis of gene expression and dependency data suggests that FHD286 sensitivity correlates with activity of genes related to DNA damage response and repair. We furthermore demonstrate an enhancing effect on tumor growth inhibition in vitro and in vivo driven by apoptosis and cell cycle arrest when combining dual FHD286 with current first and second line standards of care. Collectively, these results indicate a path forward for the development of FHD-286, a clinical stage BRM/BRG1 ATPase inhibitor, for the treatment of NSCLC patients both as a single agent treatment and in combination with current standards of care.

Citation Format: Oliver Mikse, Ammar Adam, GiNell Elliott, Dave Lahr, Victoria Amaral, Priyadharshini Viswanathan, Liv Johannessen, Hafiz Ahmad, Jessica Wan, Steven Bellon, Murphy Hentemann. FHD286 is a BRG1/BRM ATPase inhibitor showing efficacy in NSCLC models and is applicable to NSCLC patients both as a single agent treatment and in combination with current standards of care [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A079.
Abstract A089: NST-628 is a potent, fully brain-penetrant, RAS/MAPK pathway molecular glue inhibitor with efficacy in CNS tumor models
Authors: Ryan Meagan B , Li Chun , Han Yongxin , Hoeflich Klaus P , Hale Michael R , Hagel Margit
Published: December 2023

Alterations in the RAS/RAF/MEK/ERK signaling cascade are common across multiple solid tumor types and aberrant signaling of the pathway is a driver for RAS- and RAF-driven cancers. Apart from approved mutation selective inhibitors for BRAF Class I and KRAS G12C mutations, other mutations of RAS and RAF are not directly addressable by currently approved inhibitors and there is a need for inhibitors with superior efficacy, durability and tolerability for the MAPK pathway in RAS- and RAF-driven cancers. Currently approved inhibitors of the RAS-MAPK pathway also have limited central nervous system (CNS) exposure and an area of significant unmet patient need exists for patients with primary and metastatic tumors harboring RAS-MAPK pathway alterations. NST-628 has CNS permeability, with a Kpu,u >1, that is far higher than trametinib or VS-6766 (Kpu,u of 0.11 and 0.18 respectively). The predicted effective human half-life of NST-628 (~9h) is amenable to daily dosing in the clinic. In vivo, NST-628 leads to a dose-dependent inhibition of the MAPK pathway in murine brain tissue[KS1]  as measured by phospho-ERK. In an intracranial luciferase-tagged SK-MEL-2 xenograft model (NRAS Q61R), NST-628 leads to tumor regressions with a daily dosing regimen (3 mg/kg QD).  Tumor regressions as measured by bioluminescent imaging (BLI) were only seen with NST-628 (50%) in the SK-MEL-2 model, whereas trametinib and VS-6766 showed minimal efficacy due to low exposure in the CNS. In the NF-1 mutant MeWo (NF1 Q1336*) melanoma intracranial xenograft model (luciferase tagged), NST-628 led to tumor regressions at two dosing regimens (1, 3 mg/kg QD) and tumor stasis at lower doses (0.3 mg/kg) as measured by BLI. In contrast, the brain penetrant type II RAF inhibitor DAY101 (formerly TAK580) showed no efficacy in the MeWo model and resulted in paradoxical pathway hyper-activation, as determined by enhanced PD response and tumor growth. Only NST-628 effectively inhibited MAPK signaling as measured by DUSP6 transcript in a time- and dose-dependent manner. Overall, survival of mice harboring MeWo intracranial tumors was also significantly increased versus vehicle control and DAY101 treatment groups. These data collectively support best-in-class potential of NST-628 and initiation of clinical trials in patients with solid tumors harboring RAS- and RAF-mutations, including those presenting clinically with brain metastases and primary intracranial tumors.

Citation Format: Meagan B Ryan, Chun Li, Yongxin Han, Klaus P Hoeflich, Michael R Hale, Margit Hagel. NST-628 is a potent, fully brain-penetrant, RAS/MAPK pathway molecular glue inhibitor with efficacy in CNS tumor models [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A089.
Abstract A098: HBI-2376, HUYABIO clinical stage SHP2 inhibitor, displays efficacy signal in patients with KRAS mutations in early clinical studies
Authors: Shojaei Farbod , Ricono Jill M , Fang Che , Xing Yan , Ning John , Lee Gloria , Gillings Mireille
Published: December 2023

SHP2 is an attractive target in oncology drug development due to its dual roles in tumor progression: i) direct growth and proliferation of cancer cells; ii) acting as an immune checkpoint molecule to suppress tumor immunity. HBI-2376 is an orally bioavailable selective SHP2 inhibitor developed for the treatment of solid tumors harboring KRAS or EGFR mutations, such as non-small cell lung cancer (NSCLC), colorectal cancer (CRC) and pancreatic cancer (PAC). Additionally, HBI-2376 has the potential to suppress tumor growth via induction of the activity of immune infiltrating cells in the TME (tumor microenvironment). Preclinical studies showed greater potency of HBI-2376 to inhibit cell proliferation in a variety of cancer cell lines compared to other SHP2 inhibitors such as TNO-155 (Novartis AG) and RMC-4550 (Revolution Medicines). Additionally, HBI-2376 was found to be more efficacious in tumor growth inhibition than TNO-155 and RMC-4550 in NCI-H1975L858R_T790M_C797S osimertinib-resistant cell line and other tumor models. Furthermore, Western blotting showed inhibition of p-ERK and DUSP6 in HBI-2376 treated cells indicative of a biomarker of response to the therapy. In addition to xenograft models, we tested efficacy of HBI-2376 as single agent or in combination with anti-PD-1 mAb in MC38 CRC syngeneic model. HBI-2376 showed greater efficacy in tumor growth inhibition vs. TNO-155 and RMC-4550 both as single agent and in combination therapy. IHC data showed reduction in infiltration of M2 macrophages (F4/80+Arg1+) in the HBI-2376 treated tumors suggesting a mechanism of action for its greater efficacy in MC38 model. HBI-2376 is currently being tested in Phase 1 clinical trials. Consistent with preclinical studies, we have observed efficacy signals in early cohorts in two patients with partial responses (PR). Both patients were harboring KRAS mutations beyond KRAS G12C. Additionally, HBI-2376 was well tolerated with no SAEs or DLTs in the first four cohorts indicative of a good tolerability in cancer patients. Analysis of pERK in PBMCs isolated from the treated patients showed dose dependent inhibition in pERK further supporting a mechanism driven response to HBI-2376. Therefore, the preclinical and clinical data may represent HBI-2376 as the best-in-class SHP2 inhibitor.

Citation Format: Farbod Shojaei, Jill M Ricono, Che Fang, Yan Xing, John Ning, Gloria Lee, Mireille Gillings. HBI-2376, HUYABIO clinical stage SHP2 inhibitor, displays efficacy signal in patients with KRAS mutations in early clinical studies [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A098.
Abstract A099: Best in class, potent, SOS1 inhibitors demonstrate single agent activity in preclinical models of KRAS driven tumors
Authors: Lakhavath Balakrishna , Pothuganti Manoj , Kummari Githavani , Pathange Hemasankar , Sekar Rajan , Viswanadha Srikant
Published: December 2023

KRAS is an oncogene implicated in a wide variety of tumors (~21% of solid tumors harbor KRAS mutations). Interaction of KRAS with its predominant Guanine Exchange Factor (GEF), SOS1, is crucial for activation of KRAS and further KRAS mediated oncogenic signaling. Thus, pharmacological inhibition of SOS1 is expected to be effective in treating RAS driven cancers. Biochemical potency of compounds was confirmed in a SOS1 mediated FRET based guanine exchange factor (GEF) assay. Multiple potent SOS1 inhibitors were identified with potencies in the low double digit nanomolar range. Selectivity was determined against SOS2 and a host of other kinases in the RAS/RAF/MEK or RAS/PI3K/AKT pathways. Anti-proliferative activity across a panel of WT and mutant KRAS cell lines revealed IC50 between 50-200 nM with synergism observed when combined with MAPK pathway inhibitors as well as the KRAS G12C inhibitor, Sotorasib, in a 3D anchorage independent assay format. Correspondingly, significant reduction in PD biomarkers (>50% at 100 nM), pERK and pAKT, was demonstrated in KRAS mutant as well as EGFR/PI3K mutant cell lines. PK-PD correlation was also established in a tumor bearing mice model, with dose-dependent reduction of both pERK and pAKT. Compounds demonstrated excellent ADME properties with significant tissue distribution upon oral dosing. One of the lead molecules demonstrated significant single agent efficacy with growth inhibition of approximately 70% in a KRAS G12C bearing NSCLC xenograft model. No adverse signs including reduction in food intake or body weight loss were reported during the study period. IND-enabling studies are currently ongoing for the lead compounds.

Citation Format: Balakrishna Lakhavath, Manoj Pothuganti, Githavani Kummari, Hemasankar Pathange, Rajan Sekar, Srikant Viswanadha. Best in class, potent, SOS1 inhibitors demonstrate single agent activity in preclinical models of KRAS driven tumors [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A099.
Abstract A131: Cumulative burden of fatty liver and kidney cancer in young-aged men: A national population-based study
Authors: Lee Hee Yeon , Han Kyung-Do , Kwon Hyuk-Sang
Published: December 2023

Introduction Kidney cancer is considered as a metabolic disease due to its association with metabolic problems including obesity and metabolic syndrome. The association of kidney cancer with metabolic syndrome components and higher association in young-aged male population has been reported. Recent trends show an increasing incidence of kidney cancer, coupled with rising costs due to the development of new drugs. In light of these factors, identifying modifiable factors becomes a meaningful endeavor from a public healthcare perspective. The aim of this study was to determine the association of kidney cancer with cumulative burden of fatty liver in young-aged men using fatty liver index (FLI). Methods Korean National Health Insurance Service database were used. A total of 1,007,906 men aged ≥ 20 years and < 40 years who received four consecutive check-ups from January 2009 to December 2012, excluding those with missing data, history of malignancy, and those who died or diagnosed with cancer within one year after their last check-up were analyzed. They were followed up to the diagnosis of kidney cancer or death until December 2020. Non-alcoholic fatty liver disease (NAFLD) was defined as FLI ≥ 60. Results With increasing cumulative frequency of FLI >60, there was a noticeable rise in the proportion of subjects who were current smokers and heavy drinkers. Furthermore, BMI, WC, glucose and lipid profiles (excluding HDL) demonstrated an upward trend. HR in subjects with mean 30 ≤ FLI < 60 was 1.447 (95% CI 1.26-1.853) and 2.391 (95% CI 1.986-2.879) in those with mean FLI ≥ 60. As the cumulative frequency increased, HR for kidney cancer increased. A group that initially had FLI ≥ 60, and FLI decreased below 60 at the last check-up showed lower HR compared to a group that had FLI ≥ 60 on both measurements (HR 1.374 vs. 2.430).  Furthermore, this HR was lower than the group that had FLI < 60 initially but increased to ≥ 60  (1.374 vs. 1.653). Conclusion This study showed the association between cumulative burden of fatty liver using FLI and kidney cancer in young-aged men. As the cumulative burden of NAFLD increased, the incidence of kidney cancer increased.  In the cases with initial high FLI (≥ 60) and final low FLI (< 60) showed lower HR compared to the cases with high FLI at initial and final measurements. 

Citation Format: Hee Yeon Lee, Kyung-Do Han, Hyuk-Sang Kwon. Cumulative burden of fatty liver and kidney cancer in young-aged men: A national population-based study [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A131.
Abstract A136: Inferring therapeutic vulnerability within tumors through integration of pan-cancer cell line and single-cell transcriptomic profiles
Authors: Zhang Weijie , Maeser Danielle , Lee Adam , Huang Yingbo , Gruener Robert F , Jena Sampreeti , Patel Anand G , Huang R. Stephanie
Published: December 2023

Single-cell sequencing techniques have greatly advanced our current understanding of intratumoral heterogeneity through identifying tumor subpopulations with distinct biologies and therapeutic responses. However, translating biological differences into treatment strategies is challenging, as we still lack tools to facilitate efficient drug discovery that tackles heterogeneous tumors. One key step in development of such approaches centers around accurate prediction of drug response at the single-cell level to offer therapeutic options to specific cell subpopulations. Here, we present a transparent computational framework (nicknamed scIDUC) to predict therapeutic efficacies on an individual-cell basis by integrating single-cell transcriptomic profiles with large, data-rich pan-cancer cell line screening datasets. Our method detects shared expression patterns between the two data sources and utilizes such information to project cellular drug response. This method achieves high accuracy, with predicted sensitivities easily able to separate cells into their true cellular drug resistance status as measured by effect size (Cohen's d > 1.0); this holds when using single-cell RNA-seq from both cell line and in vivo models. More importantly, we examine our method’s utility with three distinct prospective tests, and in each our predicted results are accurate and mirrored biological expectations. In the first two tests, we investigated predicting drugs for cell subpopulations that are resistant to standard-of-care (SOC) therapies due to intrinsic resistance or effects of tumor microenvironments. In both, our results showed high consistency with results from the original studies. In the third test, we generated SOC therapy resistant cell lines, used scIDUC to identify drugs predicted effective on the resistant line, and validated the predictions with in vitro experiments. Together, scIDUC quickly and directly translates scRNA-seq data into meaningful cellular drug response for individual cells, displaying the potential to be used by researchers as a first-line tool for nuanced and heterogeneity-aware drug discovery.

Citation Format: Weijie Zhang, Danielle Maeser, Adam Lee, Yingbo Huang, Robert F Gruener, Sampreeti Jena, Anand G Patel, R. Stephanie Huang. Inferring therapeutic vulnerability within tumors through integration of pan-cancer cell line and single-cell transcriptomic profiles [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A136.
Abstract A138: Preclinical pharmacokinetic-pharmacodynamic modeling guides setting of the human starting dose of DS-1471a, a novel anti-CD147 antibody
Authors: Yokoyama Miki , Oitate Masataka , Sakurai Ken , Iguchi Takuma , Damme Katja , Nanai Kayoko , Fukuchi Keisuke
Published: December 2023

Background: CD147 is a member of the immunoglobulin superfamily that is highly expressed in various cancers and is involved in growth, metastasis, and activation of inflammatory signals in cancer cells. DS-1471a is a novel humanized anti-human CD147 monoclonal antibody under development as an anticancer agent. In this study, pharmacokinetic (PK)-pharmacodynamic (PD) analysis of DS-1471a was conducted to set the dose and dosing regimen for first-in-human (FIH) study. The safety profile of DS-1471a was also assessed in cynomolgus monkeys.

Methods: Preclinical pharmacological study was conducted in xenograft model mice transplanted with MIA PaCa-2 cells by intravenous (IV) administration of DS-1471a, and receptor occupancy (RO) of CD147 in tumor was measured. Monkey PK and RO in blood were measured after the IV administration of DS-1471a. The expression levels of CD147 in hepatocytes and erythrocytes in monkeys and humans were measured to estimate the species differences. Using monkey PK, RO, and CD147 expression data, a minimal physiologically based pharmacokinetic (PBPK) model was constructed. Subsequently, PK and RO profiles in humans were predicted by replacing physiological parameters and CD147 expression levels with human values. For the safety assessment, DS-1471a was intravenously administered to monkeys every 2 weeks for 1 month.

Results: In xenograft model mice, DS-1471a exhibited partial and potent antitumor activity at 1 and 3 mg/kg, respectively. At those doses, RO in tumor the day after dosing was 18% (1 mg/kg) and 89% (3 mg/kg). In monkeys, non-linear PK was observed, owing to high expression of CD147 in erythrocytes, resulting in target-mediated drug disposition. Compared to the findings in monkeys, the human expression level of CD147 was lower in erythrocytes and higher in hepatocytes. PBPK modeling, incorporating species differences in CD147 expression levels, projected that RO in tumor after the IV administration of DS-1471a to humans would reach 20%, 50%, and 90% at 2 day after injection at 0.3, 1, and 3 mg/kg, respectively. In the monkey toxicity study, DS-1471a was well tolerated at up to 158.4 mg/kg (human equivalent dose of 51 mg/kg).

Conclusions: Based on preclinical PK-PD analysis and safety study of DS-1471a, the starting dose in FIH study was set to 1 mg/kg as a safe and efficacious dose.

Citation Format: Miki Yokoyama, Masataka Oitate, Ken Sakurai, Takuma Iguchi, Katja Damme, Kayoko Nanai, Keisuke Fukuchi. Preclinical pharmacokinetic-pharmacodynamic modeling guides setting of the human starting dose of DS-1471a, a novel anti-CD147 antibody [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A138.
Abstract A153: ALK proximitome reveal SLC3A2, part of the polyamine transporter, as a membrane interaction partner with growth promoting ability
Authors: Hallberg Bengt , Lai Wei-Yun , Chuang Tzu-Po , Palmer Ruth
Published: December 2023

Solute Carrier Family 3, Member 2 (SLC3A2 or 4F2hc) was identified as a potential ALK interacting partner in a BioID-proximity labeling screen in neuroblastoma cells. SLC3A2 is a multifunctional glycoprotein that mediates integrin-dependent signaling, acts as a trafficking chaperone for amino acid transporters, and regulates polyamine transportation. However, the functional consequences of an interaction between SLC3A2 and ALK are unclear. In this work we confirm that endogenous SLC3A2 and ALK interact in NB cells. This SLC3A2:ALK interaction is abrogated upon treatment with the ALK inhibitor lorlatinib. In agreement, stimulation of ALK with ALKAL2 ligand, resulted in SLC3A2 protein levels, which was decreased upon lorlatinib treatment. We show here that loss of ALK activity leads to suppressed SLC3A2 expression and reduced SLC3A2 protein stability in a panel of neuroblastoma cells. Solute Carrier Family 7 Member 3 (SLC7A5, also known as LAT1) is an amino acid transporter that forms a stable complex with SLC3A2 for localization at the plasma membrane, where they function. This complex is lost upon knockdown of SLC3A2, with concomitant inhibition of NB cell growth. To investigate the therapeutic potential of SLC3A2/SLC7A5 complex targeting, we performed monotreatment of NB cells with AMXT-1501 (a polyamine transport inhibitor) only showed a moderate effect in NB cells. In contrast, a combined  lorlatinib/AMXT-1501 treatment resulted in synergistic inhibition of cell growth in ALK-driven primary-cultured mouse neuroblastoma cell lines. Taken together, our results show that SLC3A2 protein stability and its interaction with ALK depend on ALK signaling in ALK-driven neuroblastoma. The synergistic effect of combined ALK and polyamine transport inhibitors suggests a potential therapeutic option for ALK-driven neuroblastoma.

Citation Format: Bengt Hallberg, Wei-Yun Lai, Tzu-Po Chuang, Ruth Palmer. ALK proximitome reveal SLC3A2, part of the polyamine transporter, as a membrane interaction partner with growth promoting ability [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A153.
Abstract A154: PDLIM2 is required for regulating inflammation and cellular redox levels in mouse colorectal epithelial model
Authors: Ward Stephanie , Melgar Silvia , Cox Orla T , Morelli Raffaella , O'Connor Rosemary
Published: December 2023

PDLIM2 is a cytoskeletal and nuclear protein present in hematopoietic and epithelial cells that regulates cell differentiation and the protein stability of transcription factors including NF-κB, STATs, and β-catenin (1). PDLIM2 expression is repressed in certain cancers, but is highly expressed in cancer cells that exhibit a metastatic phenotype (2). PDLIM2 is also highly expressed in the tumour microenvironment of breast cancer, particularly in the pro-wound healing M2 macrophage population. In line with this, bone-marrow derived macrophages from PDIM2 knockout (KO) mice have impaired differentiation towards the M2 macrophage phenotype (3).  Here we investigated the effects of PDLIM2 loss in epithelial and immune cells in a colitis-associated cancer model using PDLIM2 wild type (WT) and KO mice exposed to dextran sodium sulphate (DSS) and azoxymethane (AOM). We observed that PDLIM2 KO mice exhibited a strong response to DSS alone. Colon tissue from DSS-treated mice exhibited impaired epithelium repair, sustained inflammation, and low levels of wound-healing M2 macrophages compared to WT tissues. To address the molecular mechanisms associated with PDLIM2 KO in colon epithelial cells, we suppressed or knocked out PDLIM2 in the Caco2 cell line. Caco-2 cells lacking PDLIM2 displayed dysregulated adhesion signalling, which was accompanied by elevated levels of reactive oxygen species (ROS), mitochondrial dysfunction and increased dependence on glycolysis.  Overall, we conclude that PDLIM2 is required for the resolution of inflammation and to promote intestinal epithelial repair through regulating cell adhesion signalling and cellular redox balance. 1. Guo ZS, Qu Z. PDLIM2: Signaling pathways and functions in cancer suppression and host immunity. Biochim Biophys Acta Rev Cancer. 2021 Dec;1876(2):188630. doi: 10.1016/j.bbcan.2021.188630.  2. Cox OT, Edmunds SJ, Simon-Keller K, Li B, Moran B, Buckley NE, Bustamante-Garrido M, Healy N, O'Flanagan CH, Gallagher WM, Kennedy RD, Bernards R, Caldas C, Chin SF, Marx A, O'Connor R. PDLIM2 Is a Marker of Adhesion and β-Catenin Activity in Triple-Negative Breast Cancer. Cancer Res. 2019 May 15;79(10):2619-2633. doi: 10.1158/0008-5472.CAN-18-2787.   3. Cox OT, O'Sullivan N, Tresse E, Ward S, Buckley N, O'Connor R. PDLIM2 is highly expressed in Breast Cancer tumour-associated macrophages and is required for M2 macrophage polarization. Front Oncol. 2022 Nov 30;12:1028959. doi: 10.3389/fonc.2022.1028959.

Citation Format: Stephanie Ward, Silvia Melgar, Orla T Cox, Raffaella Morelli, Rosemary O'Connor. PDLIM2 is required for regulating inflammation and cellular redox levels in mouse colorectal epithelial model [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A154.
Abstract A155: The TGF-β/BMP signaling cascades induce opposing fates in breast cancer and medulloblastoma through chromatin and cell cycle modulation
Authors: Ali Mohamad Moustafa , Ohata Yae , Heldin Carl-Henrik , Moustakas Aristidis
Published: December 2023

The transforming growth factor β (TGF-β) superfamily comprises more than 30 structurally-related proteins, including TGF-β1, TGF-β2, TGF-β3 and bone morphogenesis proteins (BMPs). TGF-β/BMP ligands bind to their respective kinase receptors culminating paradoxically in either tumor-promoting or suppressing outcomes. The former fate is attributed mainly to the induction of the epithelial-mesenchymal transition and microenvironmental remodeling by TGF-β/BMP, whereas the latter outcome is a consequence of various antiproliferative mechanisms. To gain a broader perspective, we studied the fates of different cancer models, including breast cancer and medulloblastoma, either challenged with TGF-β/BMP ligands or depleted of essential transcription factors. In 3D cultures, TGF-β stimulation enhances the genome-wide chromatin accessibility of normal-like and transformed breast epithelial models. Concomitantly, we observed a dramatic increase in accessible regions mapped to transcription start sites and, to less extent, at annotated enhancers. The corresponding transcriptome-wide analyses indicated positive modulation of DNA replication and repair associated with resistance to cyclin-dependent kinase inhibition in breast cancer organoids. Nevertheless, such TGF-β-induced remodeling was partly dependent on the activity of the transcription factor SOX4. Further implementation of whole transcriptome spatial profiling in medulloblastoma PDX tissues identified several cell cycle-related pathways and chromatin remodeling terms that negatively associate with activated BMP signaling. In this context, the stem cell capacity of medulloblastoma spheroids was significantly inhibited by BMP stimulation in vitro. Moreover, we uncovered a regulatory network that maintains the transcriptional activation of BMP receptor type I (ACVR1) through the LHX2 transcription factor that maintains an active chromatin landscape at the ACVR1 proximal promoter. The depletion of LHX2 resulted in reduced tumor formation capacity of medulloblastoma in vivo. Taken together, our extensive studies across multiple model systems suggest that the TGF-β superfamily remodels the chromatin state to alter the cell cycle progression and, subsequently, affect cancer stemness and cell fate.

Citation Format: Mohamad Moustafa Ali, Yae Ohata, Carl-Henrik Heldin, Aristidis Moustakas. The TGF-β/BMP signaling cascades induce opposing fates in breast cancer and medulloblastoma through chromatin and cell cycle modulation [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A155.
Abstract A158: Rational development of synergistic therapies alongside BMI1 Inhibition for group 3 medulloblastoma
Authors: Bakhshinyan David , Custers Stefan , Adile Ashley , Brown Kevin , Qazi Maleeha , Chokshi Chirayu , Escudero Laura , Suk Yujin , Venugopal Chitra , Moffat Jason , Singh Sheila
Published: December 2023

Medulloblastoma (MB) is the most common malignant pediatric brain tumor. Of its four distinct molecular subgroups, Group 3 MBs are associated with increased risk of recurrence, metastasis and overall poor patient outcome. In recent years, small molecule inhibitors that decrease BMI1 expression have shown to be efficacious against several types of malignant tumors including pediatric MB. Although in vivo studies provide a promising proof-of-concept for the therapeutic targeting of BMI1 in Group 3 MB, mice that receive treatment eventually succumb to their disease. These results suggest that additional mechanisms may underlie the maintenance of MB and underscore the main obstacle in treating a constantly evolving tumor. After initial preclinical validation of BMI1 inhibitor PTC-596, DNA barcoding clonal tracking technology was leveraged to profile in vivo clonal dynamics of Group 3 MB in response to the established chemoradiotherapy regimen alone and in combination with PTC-596. Comparison of clonal composition of tumors extracted from brains and spines post-treatment revealed the persistence of a small number of clones with the ability to escape therapy and drive subsequent tumor expansion. In order to better understand molecular susceptibilities of MB cells post-BMI1 inhibition, we undertook an in vitro genome-wide CRISPR/Cas9 screening to identify context-specific MB regulatory pathways that could be synergistically targeted along with BMI1. By comparing the results of the in vitro genome wide CRISPR/Cas9 screen to the essential genes in human neural stem cells (hNSCs), we identified several context specific regulators of mTOR, AKT and PLK1 pathways. Combination treatment alongside PTC-596 has demonstrated synergistic efficacy against MB cells with minimal toxicity to hNSCs in vitro and in preclinical in vivo studies. This study provides the foundation for clinical validation of small-molecule inhibitors synergistic with PTC-596 to improve the durability of remissions and extend survival of patients with treatment-refractory Group 3 MB.

Citation Format: David Bakhshinyan, Stefan Custers, Ashley Adile, Kevin Brown, Maleeha Qazi, Chirayu Chokshi, Laura Escudero, Yujin Suk, Chitra Venugopal, Jason Moffat, Sheila Singh. Rational development of synergistic therapies alongside BMI1 Inhibition for group 3 medulloblastoma [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A158.
Abstract A166: CRISPR-Cas9 genome editing in iPSCs for functional genetic screening
Authors: Isachenko Nadja , Hu Dongfang , Chenchik Alex , Diehl Paul , Tedesco Donato
Published: December 2023

Induced pluripotent stem cells (iPSCs) are widely used for disease modeling, drug discovery, and cell therapy development. However, iPSCs are difficult to engineer with an efficient CRISPR/Cas9 system for functional genetic screens. We used WTC11 human iPS cells to generate functionally validated, lentiviral-transduced Cas9 lines, characterized by high gene-editing activity and sustained potential to differentiate. Flow cytometry was used to check for the expression of pluripotency markers Oct3/4, TRA1-60, and SSEA-4 in the iPS-Cas9 cells, and their actual pluripotency was then confirmed by testing the ability to differentiate into the three germ layers: ectoderm, mesoderm, and endoderm. Furthermore, genome-wide transcriptome analysis was performed to confirm the expression of germ layer-specific markers in the differentiated cells. Sustained Cas9 activity was confirmed in the differentiated cells. This study provides proof-of-principle that patient-derived iPSCs can be used to enable CRISPR/Cas9 functional genetic screening technology in reconstituted patient-specific tissues or disease models.

Citation Format: Nadja Isachenko, Dongfang Hu, Alex Chenchik, Paul Diehl, Donato Tedesco. CRISPR-Cas9 genome editing in iPSCs for functional genetic screening [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr A166.