Journal: Advanced Science

Article Title: A Dual‐Membrane Biomimetic Nanoplatform Enables Triple‐Modal Therapy Against SARS‐CoV‐2 Through Viral Decoy, Inflammation Neutralizing, and Intracellular RNAi

doi: 10.1002/advs.77581

Figure Lengend Snippet: Schematic design and proposed therapeutic mechanism of the dual‐membrane biomimetic nanoplatform [A&T]MLN. (A) Fabrication process of [A&T]MLN. Cell membranes isolated from ACE2‐overexpressing HEK293T cells and THP‐1‐derived macrophages are fused and subsequently coated onto siRNA‐loaded lipid nanoparticles (prepared by thin‐film hydration) to generate the final nanoconstruct. (B) Proposed triple‐modal therapeutic action against SARS‐CoV‐2 infection: (i) neutralization of free virions via high‐affinity binding of surface‐displayed ACE2; (ii) attenuation of the cytokine storm through scavenging of key inflammatory mediators (e.g., IL‐6, TNF‐α) by macrophage‐derived membrane receptors; and (iii) intracellular delivery of therapeutic siRNA to suppress viral replication.

Article Snippet: Male K18‐hACE2‐2A‐CreERT2 mice (6–8 weeks old; Saiye Model Biological Research Center Co., Ltd., China) were anesthetized and administered 50 μL of SARS‐CoV‐2 S1SP (RayBiotech, USA) via intratracheal instillation at a dose of 400 mg/kg, followed by an air bolus (150 μL) to promote distribution.

Techniques: Membrane, Isolation, Derivative Assay, Infection, Neutralization, Binding Assay

Journal: Advanced Science

Article Title: A Dual‐Membrane Biomimetic Nanoplatform Enables Triple‐Modal Therapy Against SARS‐CoV‐2 Through Viral Decoy, Inflammation Neutralizing, and Intracellular RNAi

doi: 10.1002/advs.77581

Figure Lengend Snippet: Molecular basis for viral interaction and cytokine neutralization. (A) Protein composition. Coomassie blue‐stained SDS‐PAGE gel of proteins extracted from [A&T]MLN and [A&T]M; LN serves as a negative control. (B) Presence of key functional proteins. Western blot analysis confirming the transfer of ACE2, IL‐6R, TNF‐R1, IL‐1R, CD116, and CD47 to [A&T]MLN and [A&T]M; LN serves as a negative control. (C) Protein secondary structure integrity. Far‐UV circular dichroism (CD) spectra of proteins extracted from [A&T]MLN, THP‐1 membrane, [A&T]M, AMLN, and TMLN. (D) Binding kinetics to SARS‐CoV‐2 spike protein. Biolayer interferometry (BLI) sensorgrams of serially diluted [A&T]MLN (62.5–1000 n m ) binding to immobilized spike S1+S2 ECD. Red lines represent the global fit to a 1:1 binding model ( R 2 = 0.99). (E, F) Virus‐induced changes in nanoparticle properties. Hydrodynamic diameter (E) and zeta potential (F) of [A&T]MLN after 2 h incubation with authentic wild‐type (WT) or Omicron BA.5 SARS‐CoV‐2. Data are presented as mean ± SD ( n = 3). Statistical significance was determined by one‐way ANOVA. ns, not significant; ** P < 0.01; *** P < 0.001; **** P < 0.0001. (G, H) Visualization of virus‐nanoparticle complexes. Representative TEM images of [A&T]MLN bound to authentic SARS‐CoV‐2 WT (G) or Omicron BA.5 (H). Scale bars, 100 nm. (I) In vitro cytokine neutralization. Residual levels of human recombinant IL‐6, IL‐1β, TNF‐α, and GM‐CSF after incubation with AMLN, TMLN, or [A&T]MLN (0–10 µg/mL), quantified by ELISA. Data are presented as mean ± SD ( n = 3). Statistical significance was determined by one‐way ANOVA. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Male K18‐hACE2‐2A‐CreERT2 mice (6–8 weeks old; Saiye Model Biological Research Center Co., Ltd., China) were anesthetized and administered 50 μL of SARS‐CoV‐2 S1SP (RayBiotech, USA) via intratracheal instillation at a dose of 400 mg/kg, followed by an air bolus (150 μL) to promote distribution.

Techniques: Neutralization, Staining, SDS Page, Negative Control, Functional Assay, Western Blot, Circular Dichroism, Membrane, Binding Assay, Virus, Zeta Potential Analyzer, Incubation, In Vitro, Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: A Dual‐Membrane Biomimetic Nanoplatform Enables Triple‐Modal Therapy Against SARS‐CoV‐2 Through Viral Decoy, Inflammation Neutralizing, and Intracellular RNAi

doi: 10.1002/advs.77581

Figure Lengend Snippet: Broad‐spectrum antiviral activity of [A&T]MLN against SARS‐CoV‐2. (A) Dose‐response neutralization of pseudotyped SARS‐CoV‐2 variants. Neutralization activity of [A&T]MLN against a panel of pseudoviruses (WT, Alpha, Beta, Gamma, Delta, Omicron BA.4/5) was measured by a luciferase‐reporter assay. Data are presented as mean ± SD ( n = 3). (B) Contribution of the ACE2 membrane to neutralization potency. Neutralization of WT and Omicron BA.4/5 pseudoviruses by [A&T]MLN was compared to that of nanoparticles lacking the ACE2 membrane. Data are presented as mean ± SD ( n = 3). Statistical significance was determined by one‐way ANOVA. ns, not significant; * P < 0.05; **** P < 0.0001. (C) Schematic illustration of the authentic virus neutralization assay. (D) Neutralization of authentic SARS‐CoV‐2 variants. Vero E6 cells pretreated with [A&T]MLN (19.5–78 µg/mL) were infected with WT, Beta, or Omicron BA.5 viruses (MOI = 0.01). Viral titers were determined by TCID 50 . Data are presented as mean ± SD ( n = 3).

Article Snippet: Male K18‐hACE2‐2A‐CreERT2 mice (6–8 weeks old; Saiye Model Biological Research Center Co., Ltd., China) were anesthetized and administered 50 μL of SARS‐CoV‐2 S1SP (RayBiotech, USA) via intratracheal instillation at a dose of 400 mg/kg, followed by an air bolus (150 μL) to promote distribution.

Techniques: Activity Assay, Neutralization, Luciferase, Reporter Assay, Membrane, Virus, Infection