A biomimetic dual-targeting nanoplatform enhancing pyroptosis and chemosensitization for precision therapy of diffuse large b-cell lymphoma

Abstract

To elucidate the role of SHP-2 in regulating pyroptosis through the TXNIP‑NLRP3 inflammasome axis in diffuse large B‑cell lymphoma (DLBCL) and to evaluate the therapeutic efficacy and safety of a SHP099‑loaded B‑cell membrane–CD20 aptamer dual‑targeting biomimetic nanoplatform (SHP099@Bmem‑AptCD20). SHP-2 was knocked down using shRNA. Pyroptosis was assessed by Western blot, immunofluorescence, transmission electron microscopy, Annexin V/PI staining, FAM-FLICA caspase-1 assays, and apoptosis-marker analysis. Protein interactions were analyzed by co-immunoprecipitation. The nanoplatform was characterized for particle size, membrane orientation, AptCD20 density, serum stability, drug release, cellular uptake, CD20-dependent specificity, and normal B-cell safety. Pharmacokinetics, tissue distribution, antitumor activity, and preliminary safety were evaluated in NOD/SCID xenograft models. SHP-2 downregulation suppressed proliferation and invasion while inducing pyroptosis hallmarks, including GSDMD-N and cleaved caspase-1 upregulation and increased LDH, IL-1β, and IL-18. Mechanistically, SHP-2 interacted with TXNIP and limited TXNIP-NLRP3 association, whereas NLRP3 inhibition or TXNIP interference attenuated the downstream pyroptotic response. SHP-2 inhibition enhanced doxorubicin-induced pyroptosis and reduced the IC50. The SHP099@Bmem-AptCD20 nanoparticles (approximately 145 nm, PDI < 0.2) retained B-cell membrane proteins, maintained a predominantly right-side-out membrane orientation, displayed stable AptCD20 retention in serum, and showed enhanced CD20-dependent uptake. Pharmacokinetic and tissue distribution analyses indicated prolonged systemic exposure and increased tumor SHP099 accumulation, while apoptosis-marker assays supported pyroptosis-dominant mixed cell death. Normal B-cell assays and organ safety analyses indicated limited short-term toxicity under the tested conditions. SHP-2 functions as a pyroptosis suppressor by modulating the TXNIP-NLRP3 axis in DLBCL. The SHP099@Bmem-AptCD20 nanoplatform improves SHP099 delivery, supports CD20-dependent tumor targeting, and provides a rationale for further preclinical development of pyroptosis-based combination therapy.