{"doi":"10.1002/cac2.12401","title":"A nanotherapeutic strategy to target drug‐tolerant cells and overcome EGFR tyrosine kinase inhibitor resistance in lung cancer","abstract":"For patients with epidermal growth factor receptor (EGFR) mutant non-small cell lung cancer (NSCLC), EGFR tyrosine kinase inhibitors (TKIs) are used as the first-line treatment [1, 2]. Despite initial therapeutic responses, patients invariably experience disease progression due to acquired drug resistance [3]. Resistance arises, in part, because a subset of cancer cells undergoes epithelial-mesenchymal transition (EMT) and remains viable despite exposure to EGFR TKI concentrations that eliminate the bulk population [4]. The surviving cells can be re-sensitized to treatment by prolonged culture in the absence of EGFR TKIs, indicating a transient, potentially reversible, tolerance to these drugs [4]. However, these drug-tolerant cells (DTCs) may regain proliferative potential, evolve, and give rise to diverse stable mechanisms of resistance in patients [5, 6]. To address this clinical challenge, we developed a novel liposomal nanodrug, Axl-LP-VD-CTA091, to inhibit DTCs, thus targeting the origin of diverse resistance mechanisms (Figure 1A). In this formulation, the pro-differentiation agents 1,25-dihydroxyvitamin D3 (VD) and CTA091 were co-encapsulated in liposomes (LP) (Figure 1B). VD was used to suppress EMT [7] that underlies drug tolerance. CTA091 prevented catabolic inactivation of VD by 24-hydroxylase [8]. Axl aptamers [9] enabled preferential targeting of nanodrug to DTCs, which are known to have increased Axl expression [10] (see Supplementary Materials for experimental details). We prepared Axl-LP-VD-CTA091 by encapsulating VD and CTA091 in LP and then conjugated Axl aptamers on the LP surface (Supplementary Figure S1A). The size and zeta potential of Axl-LP-VD-CTA091 were 106 ± 3 nm and +30.9 ± 3.8 mV, respectively (morphology and size distribution in Supplementary Figure S1B). The encapsulation efficiency of VD and CTA091 was 75%. Axl-LP-VD-CTA091 were stable for 24 h ± serum (Supplementary Figure S1C). We evaluated the targeting and uptake of Axl-LP-VD-CTA091 in H1975OR cells that developed EMT-associated osimertinib tolerance (Supplementary Figure S2A). Axl aptamers were labeled with FITC. LP were labeled by encapsulation of Cy5-oligodeoxynucleotide (Cy5-ODN). H1975OR cells were incubated with vehicle, free Cy5-ODN, untargeted LP-Cy5-ODN or FITC-Axl-LP-Cy5-ODN. Intact FITC-Axl-LP-Cy5-ODN were taken up by H1975OR cells, as evidenced by co-localization of green and red fluorescence in the cytoplasm (Figure 1C). Flow cytometric quantitation demonstrated strong cellular uptake of FITC-Axl-LP-Cy5-ODN with 816.5-fold higher uptake vs. vehicle control and 40.9-fold higher uptake vs. free Cy5-ODN (Figure 1D). However, Axl aptamers did not increase uptake beyond what was achieved with non-targeted LP. Next, we tested the prediction that Axl-LP-VD-CTA091 promoted VD signaling, decreased EMT features and improved osimertinib sensitivity in H1975OR cells. Axl-LP-VD-CTA091 induced vitamin D target gene cytochrome P450 family 24 subfamily A member 1 (CYP24A1), increased expression of epithelial marker cadherin 1 (CDH1), and decreased expression of mesenchymal marker matrix metalloproteinase-2 (MMP2) (Figure 1E). Axl-LP-VD-CTA091 was superior to comparator treatments, including free VD + CTA091 (demonstrating the importance of drug encapsulation to activity), Axl-LP-VD (indicating the importance of using CTA091 to stabilize VD) and Axl-LP containing no VD and CTA091 (showing that Axl-aptamers and empty liposomes lack therapeutic activity). Gene expression changes induced by LP-VD-CTA091 and Axl-LP-VD-CTA091 were comparable and consistent with their similar in vitro uptake. Axl-LP-VD-CTA091 induced an 8-fold increase in CDH1 transcripts and a corresponding 3.2-fold increase in E-cadherin protein (Figure 1E). Osimertinib dose-response studies were used to test EGFR TKI sensitivity. Axl-LP-VD-CTA091 re-sensitized H1975OR cells to treatment, as evidenced by a 3.5-fold reduction in the osimertinib EC50 value compared with Axl-LP (P = 0","journal":"Cancer Communications","year":2023,"id":370608,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":6,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9485,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":649843,"name":"Lingyue Yan","orcid":"0000-0002-0908-9115","position":1,"is_corresponding":false},{"id":1030933,"name":"Yafei Su","orcid":"0000-0001-6412-6559","position":2,"is_corresponding":false},{"id":496535,"name":"Laurie J. Rich","orcid":null,"position":3,"is_corresponding":false},{"id":385260,"name":"Vui King Vincent–Chong","orcid":"0000-0003-1242-7709","position":4,"is_corresponding":false},{"id":1130336,"name":"Hannah Calkins","orcid":null,"position":5,"is_corresponding":false},{"id":536330,"name":"Saraswati Pokharel","orcid":"0000-0002-4104-462X","position":6,"is_corresponding":false},{"id":1129971,"name":"Martin Petkovich","orcid":"0000-0001-7836-3264","position":7,"is_corresponding":false},{"id":368991,"name":"Mukund Seshadri","orcid":"0000-0001-8729-9532","position":8,"is_corresponding":false},{"id":457417,"name":"Yun Wu","orcid":"0000-0002-6926-777X","position":9,"is_corresponding":false},{"id":368994,"name":"Pamela A. Hershberger","orcid":"0000-0003-0030-6520","position":10,"is_corresponding":false},{"id":370286,"name":"Tatiana Shaurova","orcid":null,"position":0,"is_corresponding":true}],"reference_count":10,"raw_metadata":null,"created_at":"2026-07-19T01:15:41.237096Z","pmid":"36691995","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}