Aug 2026· Angewandte Chemie· pp.
e6002660
· 0 citations· 44 references
Medicine
TL;DR
An acid-responsive nanoplatform using amorphous carbonated calcium phosphate nanoparticles (ACCP NPs) for effective intracellular co-delivery of TRIM21 and antibodies is disclosed, establishing ACCP NP-based TPD as a versatile and efficient platform for TRIM21/antibody-mediated degradation and a promising strategy for targeting other "undruggable" proteins.
Abstract
Targeting the intrinsically disordered oncoprotein c-Myc remains challenging due to its lack of druggable pockets, hindering small-molecule inhibitor development for decades. Antibody-based strategies utilizing TRIM21-mediated targeted protein degradation (TPD), such as TRIM-Away, offer a promising alternative for endogenous c-Myc degradation but are limited by ineffective intracellular antibody delivery and uneven endogenous TRIM21 levels in various cancer cells. To overcome these, herein we disclose an acid-responsive nanoplatform using amorphous carbonated calcium phosphate nanoparticles (ACCP NPs) for effective intracellular co-delivery of TRIM21 and antibodies. Under mild biomimetic conditions, TRIM21-antibody complexes were effectively mineralized and encapsulated in the formed nanoparticles. Following endocytic uptake, these acid-sensitive ACCP NPs disassembled in endo/lysosomes, facilitating subsequent endo/lysosomal escape and cytosolic cargo release, leading to eventual intracellular TPD. In vitro, mineralized TRIM21-antibody complexes from ACCP NPs effectively degraded various intracellular targets, particularly c-Myc, across diverse cell types, including those that are TRIM21-deficient. Specifically, m-T21-c-Myc Ab nanoparticle degraded c-Myc via the ubiquitin-proteasome system and concurrently reduced MAX levels, synergistically inhibiting c-Myc transcriptional activity. In vivo, m-T21-c-Myc Ab significantly suppressed tumor growth without major organ toxicity. This work establishes ACCP NP-based TPD as a versatile and efficient platform for TRIM21/antibody-mediated degradation and a promising strategy for targeting other "undruggable" proteins.
Microbial cell therapies hold considerable promise as programmable and versatile modalities for targeted interventions in complex biological environments. Here, we developed a living bacterial delivery platform that could leverage its endogenous metabolism to synchronize the release of surface-anchored targeted protein degradation (TPD) chimeras and the secretion of immune-modulatory nanobodies (Nbs) for enhanced antitumor efficacy. By means of metabolic labeling coupled with bioorthogonal click chemistry, transferrin (Tf)-CD24 antibody chimeras (TransCACs) were covalently displayed on the surface of nonpathogenic Escherichia coli (E. coli) K12. In parallel, this strain was equipped with a constitutive expression module for the in situ biosynthesis of PD-L1-blocking nanobodies. Capitalizing on the natural tumor tropism of bacteria, our engineered E. coli K12 achieved tumor-targeted CD24 degradation, thereby augmenting macrophage-mediated phagocytosis and synergizing with PD-L1 blockade to elicit robust tumor-specific CD8+ T cell immunity. In vivo administration of engineered microbes led to marked tumor growth inhibition in both subcutaneous breast and orthotopic hepatocellular carcinoma models, along with prolonged animal survival, driven by remodeling of the suppressive tumor microenvironment through coordinated crosstalk between M1-like macrophages and tumor-resident memory (TRM)-like CD8+ T cells. Altogether, this integrated genetic engineering and metabolic labeling of bacteria (InGeM) opens avenues for the development of next-generation microbe-based cancer immunotherapies.
Xinping Hu, Yu Chen, Meiyuan Jin et al.· Journal of the American Chem...· 0 citations
Protein–drug conjugates (PDCs) constitute a rapidly growing group of precise anticancer agents. Receptor-mediated endocytosis is a critical step in PDC action, which ensures the delivery of PDCs into the interior of cancer cells. Here, we report TriFHS-MMAE, the first multivalent PDC specifically targeting heparan sulfate proteoglycans (HSPGs) overexpressed by pancreatic cancer cells, which induces ultrafast and highly efficient aggregation-dependent endocytosis (ADE) of HSPGs. Using high-content screening with a near-kinome-wide library of inhibitors, we identified signaling pathways that govern ADE of HSPGs and discovered cascades that selectively operate in pancreatic cancer cells versus healthy cells. We show that priming cells with identified endocytic chemical modulators improves the targeting of the TriFHS-MMAE conjugate in vitro and in vivo. Overall, these findings provide insights into the interplay among signaling, endocytosis, and PDCs and support the development of specific therapies for pancreatic cancer.
Unknown authors· Journal of the American Chem...· 0 citations
Metastatic melanoma remains a critical clinical challenge due to high transcriptional plasticity and the physical barriers of the tumor microenvironment that limit the efficacy of standard therapies. In this study, we present a microfluidic-engineered poly(lactic-co-glycolic acid) nanoparticle (PLGA-NPs) platform designed to maximize therapeutic efficacy through the precise co-encapsulation of synergistic drug combinations. These monodisperse nanocarriers exhibit rapid, dose-dependent internalization across diverse patient-derived models, independent of their mutational background. Mechanistically, we demonstrate that PLGA-NPs uptake is primarily mediated by dynamin-dependent endocytosis and macropinocytosis, followed by a dynamic intracellular trafficking route that reaches a steady-state distribution within recycling endosomes, thereby enabling prolonged intracellular cargo retention. In 3D patient-derived spheroids, these nanoparticles exploit active cell-to-cell transfer consistent with transcytosis-like mechanism to penetrate the poorly accessible central core, effectively bypassing tissue-level resistance and considerably enhancing drug efficacy. In vivo biodistribution in orthotopically transplanted patient-derived xenografts confirmed highly efficient tumor targeting, while a strategic dose-modulation approach successfully optimized the tumor-to-liver accumulation ratio. Therapeutically, twice-weekly intravenous administration of trametinib-loaded NPs achieved superior tumor regression and significantly prolonged overall survival compared to daily oral administration of the free drug. Furthermore, we developed a dual-drug delivery system co-loading trametinib, MEK inhibitor, and alisertib, Aurora A kinase inhibitor. This formulation was optimized to maintain a previously established synergistic ratio which effectively overcome acquired resistance to standard-of-care therapies and induce durable tumor regression. Collectively, these findings establish the PLGA-NPs platform as a robust, clinically relevant strategy to enhance the pharmacokinetic profile of treatments for metastatic and drug-resistant melanoma.
Giulia Lovati, Pietro Milesi, Paolo Fossati et al.· Journal of Controlled Releas...· 0 citations
Neuroblastoma remains one of the most lethal pediatric solid tumors, and durable control of high-risk disease is hindered not only by inefficient tumor-selective drug delivery but also by a profoundly immunosuppressive tumor microenvironment. To address these dual barriers, we developed a chemoimmunotherapeutic vesicle-mimetic platform, DAS/CD40L-EM@DOX, by engineering CD40L-overexpressing HEK-293 T donor cells, generating extracellular vesicle mimetics through extrusion, decorating the membrane with a neuroblastoma-targeting DAS peptide, and post-loading doxorubicin. The resulting formulation preserved a nanoscale vesicular morphology, displayed CD40L on the membrane, and showed favorable particle size and zeta potential characteristics. In neuroblastoma cells, DAS decoration enhanced uptake in an α7 nicotinic acetylcholine receptor-associated manner and increased the cytotoxic and immunogenic effects of doxorubicin, as evidenced by augmented apoptosis, calreticulin exposure, and HMGB1 release. In macrophage assays, CD40L-containing vesicles shifted M2-like cells toward an M1-like phenotype, increasing CD86, TNF-α and IL-6 while decreasing CD206, IL-10 and TGF-β. Functionally, this repolarization enhanced tumor-cell phagocytosis and promoted CD8+ T-cell effector responses, including increased granzyme B, IFN-γ and IL-2 production. In tumor-bearing mice, DAS modification improved tumor accumulation and reduced off-target sequestration relative to unmodified vesicles. Therapeutically, DAS/CD40L-EM@DOX exerted the strongest inhibition of tumor growth, reduced tumor burden, prolonged survival, increased intratumoural M1-like macrophages and CD8+ T cells, decreased M2-like macrophages and regulatory T cells, and enhanced immunogenic cell-death markers in situ. Together, these findings support DAS/CD40L-EM@DOX as a dual-function vesicle-mimetic nanomedicine that couples neuroblastoma-targeted chemotherapy with macrophage reprogramming to remodel the tumor immune microenvironment and improve antitumour efficacy.
Jinkui Wang, Junyi Luo, Jiahui Li et al.· Journal of Controlled Releas...· 0 citations
Antisense oligonucleotide (ASO) therapeutics silence gene expression through RNase H-mediated mRNA degradation or steric blockade, yet their clinical efficacy is limited by inefficient intracellular trafficking, with <1% of internalized ASOs escaping endosomes to access target RNA. A deeper understanding of the cellular mechanisms governing ASO trafficking and activity is therefore critical. Here, we identify the endosomal sorting complex required for transport-I (ESCRT-I) as a previously unrecognized regulator of ASO pharmacological activity. Using human hepatocyte-derived cell models and FDA-approved ASO drugs inotersen and mipomersen, we demonstrate that suppression of specific ESCRT-I subunits, VPS23 and VPS28, markedly enhances ASO-mediated target silencing, whereas depletion of other subunits has minimal effect. Mechanistically, VPS23 functions as a central ESCRT-I component linking endosomal trafficking to intracellular glucose homeostasis. Loss of VPS23 reduces expression of the glucose transporter GLUT2, lowers intracellular glucose levels, and possibly enables ASO endosomal escape without altering cellular uptake or RNase H1-dependent activity. VPS23 suppression disrupts endosomal morphology, decreases ASO retention in late endosomes, and enhances cytosolic availability of ASOs. Notably, this regulatory effect extends beyond ASOs to small interfering RNA (siRNA) therapeutics, indicating a broader role for ESCRT-I in RNA drug biology. Together, these findings uncover a metabolic-endosomal axis controlling nucleic acid drug efficacy and provide new mechanistic insight into intracellular determinants of RNA-based therapeutics.
Le Tra Giang Nguyen, Sherouk M. Tawfik, Jing Jin et al.· Molecular Therapy: Nucleic A...· 0 citations
Background/Objectives: RNA interference (RNAi) represents a promising therapeutic approach for silencing oncogenes involved in cancer progression by utilizing small interfering RNA (siRNA). However, siRNA requires an efficient delivery system to overcome cellular uptake and endosomal escape barriers. This study aimed to evaluate a multifunctional tandem peptide, GE11-599, designed to enhance the targeted delivery of siRNA and maintain its bioactivity in glioblastoma (GBM) cells. Methods: The GE11-599 peptide, consisting of an EGFR-targeting GE11 motif and a 599 fusogenic domain, was complexed with siRNA via electrostatic interactions to form nanoparticles. We assessed nanoparticle physicochemical properties, protection of siRNA from serum and RNase degradation, and cellular uptake in two GBM cell lines (U118MG and U87MG). Mechanistic studies evaluated receptor-mediated endocytosis and the subsequent escape from endosomes. Functional assays quantified STAT3 gene silencing and downstream effects on cell migration following treatment with GE11-599–siSTAT3 complexes. Results: GE11-599 formed positively charged, monodisperse nanoparticles capable of protecting siRNA from degradation. The tandem peptide significantly enhanced cellular internalization through EGFR-mediated endocytosis and facilitated endosomal escape of siRNA. Treatment with GE11-599–siSTAT3 resulted in robust gene silencing, achieving up to an 80% reduction in STAT3 mRNA expression. Downstream functional assessment showed a 40% decrease in migration in GBM cells treated with GE11-599–siSTAT3 complexes. Conclusions: The GE11-599 tandem peptide effectively enhances cell-specific internalization and endosomal escape of siRNA in GBM cells, resulting in increased siRNA bioactivity and functional gene silencing. These findings support GE11-599 as a promising siRNA delivery platform for targeting EGFR-expressing cancers.
Jessica R. Boulos, Karen Russi, Jordan Kinnitt et al.· Pharmaceutics· 0 citations
We use cookies to run the site and, with your consent, for analytics and to show ads.
See our Cookie Policy.
