Search bioRxiv⌕ Search

bioRxiv · 10.64898/2026.05.22.727320

Interconnecting ADC Structure with Tumor Cell Biology with Multimodal Learning

Abstract

Antibody-drug conjugates (ADCs) represent a significant advancement in cancer therapy, yet their development remains constrained by high attrition rates driven by an incomplete understanding of how ADC chemical design interconnects with tumor biology. Compounding this challenge, the field has converged on a narrow set of redundant structural components, and current linker-payload systems that do not share a single mechanism of action. Existing drug response prediction frameworks cannot resolve this multidimensional complexity, relying predominantly on genomic inputs while protein-level biology is challenging to integrate. To address this, we developed a multimodal machine learning platform interconnecting ADC structural parameters with tumor cell biology across thousands of curated structure-activity datapoints, including multi-omics profiles from 1,479 human tumor cell lines and protein-level inputs from a unique model (GENCEP) that derives complete proteomic signatures. The model was validated through blinded retrospective evaluation and, critically, large coverage prospective prediction of cytotoxicity across 159 ADC-cell line combinations spanning five antigens, four mechanistically distinct linker-payload systems, and eight tumor types, most with no prior published associated ADC data. Performance surpassed industry benchmarks established for small molecule therapeutic modalities, demonstrating that protein-informed multimodal integrated framework is effective at capturing cytotoxic determinants at scale.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Mslati, H., Wilson, M., Naeinipour, M., Coulombe, G., Ezzine, M., Yuen, T., Bari, O., Singh, H., Tam, R., Sheff, J., Gentile, F., Leyton, J.. 2026-05-27. Interconnecting ADC Structure with Tumor Cell Biology with Multimodal Learning. https://doi.org/10.64898/2026.05.22.727320

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Peptide Sequence Determines the Efficacy of Single-Dose AAV-Vectorized GLP-1R Agonists in Diet-Induced Obese Mice

Background: Glucagon-like peptide-1 receptor (GLP-1R) agonists such as semaglutide produce substantial weight loss and glycemic improvement, but their long-term efficacy depends on sustained adherence to chronic dosing. Adeno-associated virus (AAV) gene therapy could, in principle, provide durable GLP-1R agonist (GLP-1RA) exposure from a single administration, but the degree of peptide engineering required for a vectorized GLP-1RA transgene to be efficacious in vivo is unknown. Methods: We generated four self-complementary AAV (scAAV) constructs encoding: native human GLP-1(7-36) (GLP-1), a DPP-IV-resistant Gly8-GLP-1(7-36) variant (Gly8), Exendin-4 (Ex4), or an Exendin-4 human IgG4 Fc fusion protein (Ex4-Fc). All four constructs were validated to secrete functional GLP-1R agonists in vitro using a conditioned-media potency assay. Subsequently, these constructs were packaged into PHP.eB capsids and administered as a single intravenous dose (5x1012 vg/kg) to diet-induced obese (DIO) mice alongside a vehicle control. Body weight, body composition, food intake, fasting plasma insulin, and oral glucose tolerance were assessed over a 3-week period. At the end of the study, tissues were harvested for assessments of plasma/liver lipids, tissue transgene transcript and peptide levels. Results: HEK293 cells transfected with the four AAV constructs all secreted active peptides/proteins as assessed by a cell-based GLP-1R activation assay, although the effective concentration of the GLP-1 constructs in the media spanned nearly four orders of magnitude 72 hours after transfection. In diet-induced obese (DIO) mice, Ex4 and Ex4-Fc vectors, but not GLP-1 or Gly8 vectors, produced significant reductions in body weight of up to 22.6% of starting body weight out to 21 days after a single dose. In addition, reductions in food intake, fat mass, and fasting insulin relative to vehicle were also observed for Ex4 and Ex4-Fc. Furthermore, Ex4-Fc improved oral glucose tolerance while reducing plasma cholesterol and liver triglycerides relative to vehicle. Although transcript levels did not differ appreciably among the four constructs in either brain or liver, measurable peptide product in plasma and brain lysate tracked the rank order of the in vitro supernatant screen, with GLP-1 and Gly8 groups indistinguishable from endogenous background. Conclusions: PHP.eB scAAV Ex4 and Ex4-Fc showed robust efficacy on multiple metabolic endpoints including weight loss and glucose tolerance. In contrast, vectorized GLP-1 and Gly8 showed no in vivo efficacy despite comparable levels of viral transcript in the brain and the liver. Vectorized GLP-1 and Gly8 also performed poorly compared to exendin-4-based constructs in vitro, suggesting that conditioned-media potency screening is a valid early filter for construct selection in vectorized-peptide AAV development. Engineering peptides to improve translation and secretion efficiency, in addition to circulating plasma stability, could further optimize vectorized GLP-1RA therapeutics.

pharmacology and toxicology↗

Preclinical Evaluation of a Novel BOC-Modified Triazoloacridinone (TRA-01AR) in Combination with 5-Fluorouracil in Colorectal Cancer

5-Fluorouracil (5-FU) remains a cornerstone of colorectal cancer (CRC) therapy, yet its clinical efficacy is limited by resistance, systemic toxicity, and unfavorable pharmacokinetics. Strategies that enhance tumor sensitivity while maintaining tolerability are therefore of considerable interest. Here, we evaluated 5-FU in combination with two triazoloacridinones, C-1305 and a novel BOC-modified derivative, TRA-01AR, using integrated cellular, transcriptomic, bioenergetic, and in vivo approaches. Both compounds induced cytotoxic effects across cancer cell models and produced overlapping but distinct transcriptional responses, primarily affecting DNA replication and cell cycle pathways. Bioenergetic profiling revealed a shift toward oxidative phosphorylation accompanied by reduced glycolysis, indicating modulation of cellular metabolism relevant to chemotherapy response. Interestingly, TRA-01AR exhibited delayed cytotoxicity relative to C-1305, although the underlying mechanism remains to be defined. In vivo, the combination of 5-FU with C-1305 significantly reduced tumor weight in HT-29 xenografts, whereas 5-FU combined with TRA-01AR did not result in a statistically significant reduction compared to 5-FU alone. TRA-01AR was associated with a more favorable systemic profile and increased intratumoral blood flow compared with C-1305. Collectively, these findings indicate that triazoloacridinones modulate transcriptional and metabolic programs relevant to the 5-FU response. While C-1305 demonstrated greater antitumor activity under the tested conditions, TRA-01AR exhibited distinct biological effects, including improved tolerability and vascular modulation. Further studies are required to clarify TRA-01AR's mechanism of action and assess its potential relevance in CRC-specific therapies.

pharmacology and toxicology↗

Molecular Mechanisms of Quizartinib Resistance in FLT3 Mutants

Quizartinib, a type II FLT3 inhibitor approved for FLT3-ITD-positive acute myeloid leukemia (AML), is frequently limited by acquired resistance, which arises in up to 40% of patients with disease progression and is driven largely by the D835Y and F691L kinase-domain mutations. To define the underlying mechanisms, we performed triplicate 200-ns all-atom molecular dynamics simulations of wild-type FLT3, FLT3-D835Y, and FLT3-F691L in complex with Quizartinib, combining conformational analysis, MM-GBSA binding free-energy calculations, and binding-pocket characterization. Both mutations destabilized the inactive DFG-out conformation required for type II inhibitor binding, with mean binding free-energy reductions of 5 kcal/mol for D835Y and 10 kcal/mol for F691L relative to wild-type. D835Y disrupted the activation-loop electrostatic network and reduced Cys694 hydrogen-bond occupancy from 85% to 45%, whereas F691L abolished key van der Waals contacts and opened a 120 cubic Angstrom hydrophobic void at the gatekeeper position. Both mutants sampled a wider range of conformations and lost 14-17% of binding-pocket volume. These predictions were corroborated by CCK-8 viability assays and Western blotting of p-FLT3 (Y591) in Ba/F3 cells, which reproduced the predicted resistance hierarchy (F691L > D835Y > WT) at the cellular level. Together, these results indicate that D835Y and F691L confer resistance through distinct electrostatic and hydrophobic mechanisms that converge on destabilization of the inactive kinase conformation, providing a structural basis for the design of next-generation FLT3 inhibitors.

pharmacology and toxicology↗