Journal: Bioactive Materials

Article Title: Dynamic feedback BacGuard anchors microbial metabolism to host symbiosis in real-time ulcerative colitis therapy

doi: 10.1016/j.bioactmat.2026.05.060

Figure Lengend Snippet: BacGuard promotes colon tissue repairment. a) Scheme of microbiota-dependent epithelial repair mechanism orchestrated by BacGuard. b) Short-chain fatty acid (SCFA) profile alterations. n = 6. c) BacGuard-induced probiotic proliferation and d) quantitative results. n = 3. e) Immunofluorescence analysis of ILC3 (ROR γt + CD3 − cells) in colon tissue. f) Flow cytometric analysis of lamina propria lymphocytes (ROR γt + ). n = 3. g) Concentration of IL-22 in MNK-3 cells. n = 3. h) Representative PAS-staining (upper panel) and MUC-2 immunohistochemistry (lower panel) images of colon tissues. n = 5. ns, not significant; ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001.

Article Snippet: For the detection of IL-22 and LPS concentrations, the Mouse IL-22 Precoated ELISA Kit (DAKEWE, China) and Mouse LPS ELISA Kit (JONLNBIO, China) were employed correspondingly.

Techniques: Immunofluorescence, Concentration Assay, Staining, Immunohistochemistry

Journal: Aging Cell

Article Title: uPAR ‐Targeting T Cell Engager Exerts Senolytic Effects in Mice and Non‐Human Primates With Serum Aminotransferase Activity as a Safety Monitor

doi: 10.1111/acel.70742

Figure Lengend Snippet: hGFD‐CD3 mediates the clearance of senescent cells by T cells in vitro. (A) A schematic diagram of hGFD‐CD3 and the negative control used in this study. (B) Semiquantitative analysis of the cell surface binding of hGFD‐CD3 to young uPAR‐HEL1 cells and uPAR+ HEL1 senescent cells via flow cytometry. (C) Semiquantitative analysis of the cell surface binding of hGFD‐CD3 and hCD3 to human CD3+ T cells via flow cytometry. (D) Dose–response study of hGFD‐CD3 mediated T cell cytotoxicity against senescent cells (E: T = 1: 1). Each data point represents the mean of a biological replicate ( n = 2). (E) Cytokine release studies using ELISA from cell supernatants in experiments where 100 ng/mL hGFD‐CD3 was applied. (F) T cell activation was assessed by measuring CD25 and CD69 levels on CD3+ T cells by flow cytometry. All experiments were performed in biological triplicate. Data are presented as mean ± SD of triplicate samples and analyzed by two‐tailed unpaired Student's t ‐test with Welch's correction (e, f). ** p ≤ 0.05; ** p ≤ 0.01; ns, not significant.

Article Snippet: The cytokine IL‐6 was measured with an ELISA kit (Proteintech, KE10007).

Techniques: In Vitro, Negative Control, Analysis, Binding Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Activation Assay, Two Tailed Test

Journal: Aging Cell

Article Title: uPAR ‐Targeting T Cell Engager Exerts Senolytic Effects in Mice and Non‐Human Primates With Serum Aminotransferase Activity as a Safety Monitor

doi: 10.1111/acel.70742

Figure Lengend Snippet: mGFD‐CD3 safely and effectively mediates T‐cell clearance of senescent cells in naturally aged mice. (A) The half‐lives of mGFD‐CD3 and mCD3 negative control in naturally aged mice were analyzed by ELISA. Each data point represents the mean of a biological replicate ( n = 2). (B) The mRNA expression levels for mouse uPAR, P16 INK4a , and P21 were detected by qPCR analysis from the indicated tissues from mice treated with mCD3 or mGFD‐CD3 on day 21 ( n = 5 mice per group). (C) Representative images of SA‐β‐Gal staining of liver and lung tissue samples collected from naturally aged mice treated with mCD3 or mGFD‐CD3 ( left ), with quantification of average staining intensities (ImageJ) ( right ) ( n = 5). Scale bar, 100 μm. (D) Images of subcutaneous adipose tissue (SAT) after SA‐β‐Gal staining ( left ) and quantification of the average staining intensities (ImageJ) ( right ). SAT was collected from mice treated with mCD3 or mGFD‐CD3 on day 21 ( n = 5 individual samples per condition). Results from mouse experiments are representative of two independent experiments. (E) Analysis of Senmayo enrichment dataset of liver tissue samples from naturally aged mice treated with mGFD‐CD3 or mCD3 as negative control ( n = 3 mice per condition). (F) Gene expression volcano plot from the analysis of liver tissue samples collected from mice treated with mCD3 or mGFD‐CD3 ( n = 3 per condition). (G) Hierarchical clustering heatmap of the senescence‐associated secretory phenotype (SASP) profiles of liver samples ( n = 3 per condition). (H) Representative images of BiTEs (green), CD3 (red), and SA‐β‐Gal staining (blue green) for liver and lung samples collected from mice treated with BiTEs 24 h post‐injection. (I) Flow cytometry results of CD3 + T cells analyzed for CD25 expression after administration of mCD3 or mGFD‐CD3 ( n = 3 mice per condition). Data is presented as mean ± SD and analyzed by one‐way ANOVA followed by Tukey's multiple comparisons test (C, D, I). * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant.

Article Snippet: The cytokine IL‐6 was measured with an ELISA kit (Proteintech, KE10007).

Techniques: Negative Control, Enzyme-linked Immunosorbent Assay, Expressing, Analysis, Staining, Tissue, Gene Expression, Injection, Flow Cytometry

Journal: Aging Cell

Article Title: uPAR ‐Targeting T Cell Engager Exerts Senolytic Effects in Mice and Non‐Human Primates With Serum Aminotransferase Activity as a Safety Monitor

doi: 10.1111/acel.70742

Figure Lengend Snippet: Elimination of senescent cells with mGFD‐CD3 ameliorated age‐related phenotypes in mice. (A) ELISA analysis for TNF‐α and IL‐1β in blood samples collected from mice treated with mGFD‐CD3 or mCD3 at 3 Months. ( n = 5 for young group; n = 3 for mCD3 group; n = 3 for mGFD‐CD3 group). (B) ( left ) H&E staining of IAT samples collected from mice described in (a). Quantification of adipocyte areas ( right ). Each data point represents an individual mouse. Scale bar, 50 μm. (C) ( left ) Masson staining of liver and lung tissues collected from the mice described in (a). Black arrows indicate positive staining. Scale bar, 50 μm. ( right ) Quantification of the fibrotic areas of liver or lung tissues using Image J. (D) PAS staining of kidney samples from mice described in (a). Scale bar, 50 μm (original) and 15 μm (magnified). Quantification was achieved using Image J. (E) TRAP and Masson staining of femur samples from mice described in (a). Black arrows indicate positive staining. Scale bar, 50 μm. Quantification was achieved with Image J. Data are presented as mean ± SD and analyzed by one‐way ANOVA followed by Tukey's multiple comparisons test (A–E). * p ≤ 0.05.

Article Snippet: The cytokine IL‐6 was measured with an ELISA kit (Proteintech, KE10007).

Techniques: Enzyme-linked Immunosorbent Assay, Analysis, Blood/Plasma/Serum Samples, Staining

Journal: Aging Cell

Article Title: uPAR ‐Targeting T Cell Engager Exerts Senolytic Effects in Mice and Non‐Human Primates With Serum Aminotransferase Activity as a Safety Monitor

doi: 10.1111/acel.70742

Figure Lengend Snippet: Assessing safety attributes of hGFD‐CD3 treatment in aged nonhuman primates. (A) Semiquantitative analysis of cell surface binding of hGFD‐CD3 to macaque T cells via flow cytometry. (B) Monitoring of liver function after administration of each different dose within a 72‐h period. (C) Schematic diagram of the experiment to investigate clearance of senescent cells from macaques, where hGFD‐CD3 was administered at a dose of 20 μg/kg every three days. (D) Liver function was evaluated throughout the injection phase. (E) Changes in cytokine levels in serum samples from three macaques before, as well as 24‐ and 72 h after the first injection of hGFD‐CD3, as determined via ELISA. Data are presented as mean ± SD of triplicate samples analyzed by two‐tailed unpaired Student's t ‐test with Welch's correction. (F) Hematological parameters were assessed throughout the injection phase. (G) Six accessible vital sign parameters were continuously monitored during the injection period. Data is presented as mean ± SD of triplicates and analyzed by two‐tailed unpaired Student's t test with Welch's correction (E). *** p ≤ 0.001.

Article Snippet: The cytokine IL‐6 was measured with an ELISA kit (Proteintech, KE10007).

Techniques: Analysis, Binding Assay, Flow Cytometry, Injection, Blood/Plasma/Serum Samples, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Aging Cell

Article Title: uPAR ‐Targeting T Cell Engager Exerts Senolytic Effects in Mice and Non‐Human Primates With Serum Aminotransferase Activity as a Safety Monitor

doi: 10.1111/acel.70742

Figure Lengend Snippet: hGFD‐CD3 clears senescent cells in aged nonhuman primates. (A) SA‐β‐Gal staining of subcutaneous adipose tissue and quantification before injection, as well as at Days 42 and 90 after the first injection. (B) Fold‐changes in mRNA expression levels of uPAR in subcutaneous adipose tissue, muscle, and skin before injection and at Days 42 and 90 after the first injection. (C) Fold‐changes in mRNA expression levels of the senescence markers P16 INK4a and P21 in subcutaneous adipose tissue, muscle, and skin before injection and at Days 42 and 90 after the first injection. (D) The expression levels of representative genes from T cell‐, NK‐, Monocyte‐, Macrophage‐, DC‐, B cell‐ and Plasma‐cell types in liver, fat, and skin tissue collected from three macaques before injection, and at Days 42 and 90 after the first injection of hGFD‐CD3, and the data of pre‐administration were normalized and compared to those of the data of day 42. ( n = 3). (E) Changes in cytokine levels in serum samples from three macaques before injection, and at Days 42 and 90 after the first injection of hGFD‐CD3, as determined via ELISA. Data is presented as mean ± SD of triplicates and analyzed by paired two‐tailed Student's t ‐test. ** p ≤ 0.01; *** p ≤ 0.001.

Article Snippet: The cytokine IL‐6 was measured with an ELISA kit (Proteintech, KE10007).

Techniques: Staining, Tissue, Injection, Expressing, Clinical Proteomics, Blood/Plasma/Serum Samples, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Systemic TREM1 ablation attenuates neuroinflammation and neurological deficits after TBI. (a) Schematic timeline of the experimental design. WT and Trem1 −/− mice were subjected to CCI or sham surgery, followed by longitudinal behavioral and histological assessments. (b, c) Analysis of the neuroinflammatory profile in the perilesional cortex at 3 days post‐TBI. (b) Representative images of inflammatory cytokines in TBI lesions detected by the Proteome Profiler Array Mouse Cytokine Array Panel A in WT and Trem1 −/− mice at 3 days post‐TBI. (c) Quantification of inflammatory cytokines in TBI lesions of WT and Trem1 −/− mice at 3 days post‐TBI or sham treatment by ELISA ( n = 5 mice per group, two‐way ANOVA followed by Tukey's multiple‐comparison test). (d) Representative immunofluorescence images of macrophages (IBA1 + , green) and the reactivation marker CD86 (red) in the TBI cortex. Trem1 ablation significantly reduced reactive polarization (scale bar, 50 µm). (e) Evaluation of neuronal apoptosis. Representative co‐staining of TUNEL (green) and NeuN (red) (left) ( n = 3 mice per group, scale bar, 100 µm). Yellow arrows indicate apoptotic neurons. (f) Assessment of white matter integrity. Representative images of Myelin Basic Protein (MBP, green) and NeuN (red) (left) and quantification of MBP fluorescence intensity (right) (scale bar, 100 µm). (g) Representative immunofluorescence images of hippocampal neurons (NeuN, red) showing preserved cellular density in Trem1 −/− mice ( n = 3 mice per group; scale bar, 100 µm). (h) Brain water content analysis at 1day and 3 d post‐TBI, indicating alleviated cerebral edema in Trem1 −/− mice ( n = 6 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (i, j) Assessment of blood‐brain barrier (BBB) permeability. (i) Representative images of Evans Blue extravasation. (j) Quantification of dye leakage ( n = 6 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (k, l) Molecular validation of tissue preservation. (k) Representative Western blots of the neuronal marker beta3‐tubulin, synaptic marker Synaptophysin (Syn), and tight junction protein Occludin. l, Densitometric quantification normalized to GAPDH ( n = 3 independent experimental replicates; two‐way ANOVA followed by Tukey's multiple‐comparison test). (m–o) Assessment of sensorimotor function. m, Latency to fall in the accelerating Rotarod test ( n = 8 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (n) Performance in the fixed‐speed (30 rpm) Rotarod test ( n = 8 mice per group, one‐way ANOVA followed by Tukey's multiple‐comparison test). o, Modified Neurological Severity Score (mNSS) trajectory ( n = 8 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (p, q) Evaluation of anxiety‐like behavior and exploratory activity in the Open Field Test (OFT). (p) Representative locomotor tracking plots. (q) Quantification of the percentage of time spent in the center zone ( n = 8 per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). Data are presented as mean ± SD. Each dot represents an individual mouse, except in l, where each dot represents an independent experimental replicate. Statistical tests are specified for each panel above. Exact p values are provided in the Source Data. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐10 ELISA kit , ABclonal , # RK00016.

Techniques: Analysis, Enzyme-linked Immunosorbent Assay, Comparison, Immunofluorescence, Marker, Staining, TUNEL Assay, Fluorescence, Permeability, Biomarker Discovery, Tissue, Preserving, Western Blot, Modification, Activity Assay

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Generative AI‐driven design and mechanistic validation of a TREM1‐targeting LYTAC degrader. (a) Schematic of the RFdiffusion‐guided pipeline for de novo TREM1‐binding peptide design. Peptide backbones were generated around the Ig‐like domain of TREM1 and subsequently screened by AlphaFold3, ZDOCK, HDOCK, and Rosetta‐based energetic analysis to identify high‐ranking candidates. (b) Predicted structural models of Pep15, Pep17, and Pep457 in complex with TREM1, showing the predicted binding interfaces. (c) Temperature‐related intensity change (TRIC) assays showing concentration‐dependent target engagement of Pep15, Pep17, and Pep457 with TREM1. Left, schematic illustration of the TRIC assay principle; right, dose‐response curves of the indicated peptides ( n = 3 independent experiments). (d) ELISA‐based binding analysis of Pep15, Pep17, and Pep457 to immobilized TREM1. Top, schematic of the ELISA workflow; bottom, quantitative binding curves ( n = 3 independent experiments). (e) Surface plasmon resonance (SPR) analysis of Pep457 binding to TREM1. Top, schematic of the SPR assay; bottom, representative sensorgrams obtained at the indicated Pep457 concentrations. (f) Structural model of the bifunctional TREM1‐LYTAC, in which Pep457 engages TREM1 and the mannose‐6‐phosphate (M6P) moiety recruits the cation‐independent mannose‐6‐phosphate receptor (CI‐M6PR). (g) Representative confocal images showing partial colocalization of TREM1‐LYTAC with lysosomes following cellular uptake. TREM1‐LYTAC, green; lysosomes, red; nuclei, blue. Arrows indicate representative colocalized puncta. Scale bars as indicated. (h) ELISA quantification of cellular TREM1 protein following treatment with vehicle, scrambled‐LYTAC, TREM1‐LYTAC, or TREM1‐LYTAC in the presence of bafilomycin A1 (BafA1) or MG132 ( n = 5 independent experiments). (i) Representative flow‐cytometric histograms showing cell‐surface TREM1 expression after the indicated treatments. (j) Trem1 mRNA expression measured by qPCR after the indicated treatments ( n = 3 independent experiments; one‐way ANOVA followed by Tukey's multiple‐comparison test). (k) Requirement of both functional modules for TREM1 reduction. Cellular TREM1 protein levels were quantified after treatment with Pep457 plus M6P, Pep457 alone, M6P alone, TREM1‐LYTAC in the presence of CI‐M6PR blockade, or conjugated TREM1‐LYTAC ( n = 5 independent experiments; one‐way ANOVA followed by Tukey's multiple‐comparison test). (l) Plasma pharmacokinetic profile of free TREM1‐LYTAC after intravenous administration. Fluorescence intensity was fitted to a monoexponential decay model, yielding an apparent half‐life of 1.13 h ( n = 3 mice). (m) Proposed mechanism of TREM1‐LYTAC‐mediated degradation. TREM1‐LYTAC bridges cell‐surface TREM1 to CI‐M6PR, thereby promoting receptor internalization and lysosomal degradation. Data are mean ± SD. Each dot represents an independent experimental replicate or an individual animal. Statistical tests are specified for each quantitative panel above. Exact p values are provided in the Source Data. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐10 ELISA kit , ABclonal , # RK00016.

Techniques: Biomarker Discovery, Binding Assay, Generated, Analysis, Concentration Assay, Drug discovery, Enzyme-linked Immunosorbent Assay, SPR Assay, Confocal, Expressing, Comparison, Functional Assay, Clinical Proteomics, Fluorescence

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Engineering and validation of a BBB‐targeted, pH‐responsive nanodevice for spatiotemporal delivery of TREM1‐LYTAC. (a) Schematic illustration of APTL‐NP fabrication. TREM1‐LYTAC was conjugated to a generation‐4 PAMAM dendrimer through a terephthalaldehyde‐derived acid‐labile linker, followed by surface functionalization with Angiopep‐2 and PS‐PEG‐NHS to enable LRP1‐directed BBB targeting and improve systemic stability. (b) Representative transmission electron microscopy images of PAMAM‐based precursor particles and APTL‐NP (Scale bars, 1 µm). (c) Dynamic light scattering profiles showing the apparent hydrodynamic size distributions of G4 PAMAM and APTL‐NP under the indicated measurement conditions. (d) Time‐dependent changes in hydrodynamic diameter and polydispersity index of APTL‐NP incubated in PBS at pH 7.4 and 37°C. (e) pH‐dependent zeta‐potential profile of APTL‐NP. (f) Nanoparticle tracking analysis showing pH‐dependent particle size and concentration distributions after incubation at pH 7.5, 7.0, or 6.5. (g) pH‐responsive release of TREM1‐binding‐competent TREM1‐LYTAC from APTL‐NP. Released free TREM1‐LYTAC was separated by ultrafiltration and quantified using a TREM1‐binding ELISA with free TREM1‐LYTAC standards ( n = 3 independent experiments). (h) Representative EdU staining images of BMECs, HT22 cells, and NHAs treated with vehicle or APTL‐NP. (i) Quantification of EdU‐positive cells in the indicated cell types( n = 6 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (j) Representative flow‐cytometric plots for apoptosis analysis in BMECs, HT22 cells, and NHAs after treatment with vehicle or APTL‐NP. (k) Quantification of apoptotic cells in the indicated cell types( n = 6 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (l) Cell viability after 24 h or 48 h exposure to the indicated concentrations of APTL‐NP ( n = 3 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (m) Plasma pharmacokinetic profile of APTL‐NP after intravenous administration. The fluorescence signal was fitted to a monoexponential decay model, yielding an apparent half‐life of 8.38 h ( n = 3 mice). (n) Representative photographs and quantification of hemolysis induced by the indicated concentrations of APTL‐NP ( n = 3 independent experiments, one‐way ANOVA followed by Tukey's multiple‐comparison test). (o, p) Plasma C3a and C5a concentrations at the indicated time points after administration of vehicle or APTL‐NP ( n = 6 mice per group, Two‐way ANOVA followed by Tukey's multiple‐comparison test). (q) Representative H&E‐stained sections of the spleen, heart, lung, liver, and kidney from vehicle‐ and APTL‐NP‐treated mice (Scale bar, 100 µm). Data are presented as mean ± SD where applicable. For in vitro assays, n denotes independent experimental replicates; for in vivo analyses, n denotes individual mice. Statistical tests are specified for each quantitative panel above. Exact p values are provided in the Source Data. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐10 ELISA kit , ABclonal , # RK00016.

Techniques: Biomarker Discovery, Derivative Assay, Stability, Transmission Assay, Electron Microscopy, Dynamic Light Scattering Assay, Incubation, Zeta Potential Analyzer, Analysis, Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Staining, Comparison, Clinical Proteomics, Fluorescence, In Vitro, In Vivo

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Key resources table.

Article Snippet: IL‐10 ELISA kit , ABclonal , # RK00016.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Systemic TREM1 ablation attenuates neuroinflammation and neurological deficits after TBI. (a) Schematic timeline of the experimental design. WT and Trem1 −/− mice were subjected to CCI or sham surgery, followed by longitudinal behavioral and histological assessments. (b, c) Analysis of the neuroinflammatory profile in the perilesional cortex at 3 days post‐TBI. (b) Representative images of inflammatory cytokines in TBI lesions detected by the Proteome Profiler Array Mouse Cytokine Array Panel A in WT and Trem1 −/− mice at 3 days post‐TBI. (c) Quantification of inflammatory cytokines in TBI lesions of WT and Trem1 −/− mice at 3 days post‐TBI or sham treatment by ELISA ( n = 5 mice per group, two‐way ANOVA followed by Tukey's multiple‐comparison test). (d) Representative immunofluorescence images of macrophages (IBA1 + , green) and the reactivation marker CD86 (red) in the TBI cortex. Trem1 ablation significantly reduced reactive polarization (scale bar, 50 µm). (e) Evaluation of neuronal apoptosis. Representative co‐staining of TUNEL (green) and NeuN (red) (left) ( n = 3 mice per group, scale bar, 100 µm). Yellow arrows indicate apoptotic neurons. (f) Assessment of white matter integrity. Representative images of Myelin Basic Protein (MBP, green) and NeuN (red) (left) and quantification of MBP fluorescence intensity (right) (scale bar, 100 µm). (g) Representative immunofluorescence images of hippocampal neurons (NeuN, red) showing preserved cellular density in Trem1 −/− mice ( n = 3 mice per group; scale bar, 100 µm). (h) Brain water content analysis at 1day and 3 d post‐TBI, indicating alleviated cerebral edema in Trem1 −/− mice ( n = 6 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (i, j) Assessment of blood‐brain barrier (BBB) permeability. (i) Representative images of Evans Blue extravasation. (j) Quantification of dye leakage ( n = 6 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (k, l) Molecular validation of tissue preservation. (k) Representative Western blots of the neuronal marker beta3‐tubulin, synaptic marker Synaptophysin (Syn), and tight junction protein Occludin. l, Densitometric quantification normalized to GAPDH ( n = 3 independent experimental replicates; two‐way ANOVA followed by Tukey's multiple‐comparison test). (m–o) Assessment of sensorimotor function. m, Latency to fall in the accelerating Rotarod test ( n = 8 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (n) Performance in the fixed‐speed (30 rpm) Rotarod test ( n = 8 mice per group, one‐way ANOVA followed by Tukey's multiple‐comparison test). o, Modified Neurological Severity Score (mNSS) trajectory ( n = 8 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (p, q) Evaluation of anxiety‐like behavior and exploratory activity in the Open Field Test (OFT). (p) Representative locomotor tracking plots. (q) Quantification of the percentage of time spent in the center zone ( n = 8 per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). Data are presented as mean ± SD. Each dot represents an individual mouse, except in l, where each dot represents an independent experimental replicate. Statistical tests are specified for each panel above. Exact p values are provided in the Source Data. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐1β ELISA kit , ABclonal , # RK00006.

Techniques: Analysis, Enzyme-linked Immunosorbent Assay, Comparison, Immunofluorescence, Marker, Staining, TUNEL Assay, Fluorescence, Permeability, Biomarker Discovery, Tissue, Preserving, Western Blot, Modification, Activity Assay

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Generative AI‐driven design and mechanistic validation of a TREM1‐targeting LYTAC degrader. (a) Schematic of the RFdiffusion‐guided pipeline for de novo TREM1‐binding peptide design. Peptide backbones were generated around the Ig‐like domain of TREM1 and subsequently screened by AlphaFold3, ZDOCK, HDOCK, and Rosetta‐based energetic analysis to identify high‐ranking candidates. (b) Predicted structural models of Pep15, Pep17, and Pep457 in complex with TREM1, showing the predicted binding interfaces. (c) Temperature‐related intensity change (TRIC) assays showing concentration‐dependent target engagement of Pep15, Pep17, and Pep457 with TREM1. Left, schematic illustration of the TRIC assay principle; right, dose‐response curves of the indicated peptides ( n = 3 independent experiments). (d) ELISA‐based binding analysis of Pep15, Pep17, and Pep457 to immobilized TREM1. Top, schematic of the ELISA workflow; bottom, quantitative binding curves ( n = 3 independent experiments). (e) Surface plasmon resonance (SPR) analysis of Pep457 binding to TREM1. Top, schematic of the SPR assay; bottom, representative sensorgrams obtained at the indicated Pep457 concentrations. (f) Structural model of the bifunctional TREM1‐LYTAC, in which Pep457 engages TREM1 and the mannose‐6‐phosphate (M6P) moiety recruits the cation‐independent mannose‐6‐phosphate receptor (CI‐M6PR). (g) Representative confocal images showing partial colocalization of TREM1‐LYTAC with lysosomes following cellular uptake. TREM1‐LYTAC, green; lysosomes, red; nuclei, blue. Arrows indicate representative colocalized puncta. Scale bars as indicated. (h) ELISA quantification of cellular TREM1 protein following treatment with vehicle, scrambled‐LYTAC, TREM1‐LYTAC, or TREM1‐LYTAC in the presence of bafilomycin A1 (BafA1) or MG132 ( n = 5 independent experiments). (i) Representative flow‐cytometric histograms showing cell‐surface TREM1 expression after the indicated treatments. (j) Trem1 mRNA expression measured by qPCR after the indicated treatments ( n = 3 independent experiments; one‐way ANOVA followed by Tukey's multiple‐comparison test). (k) Requirement of both functional modules for TREM1 reduction. Cellular TREM1 protein levels were quantified after treatment with Pep457 plus M6P, Pep457 alone, M6P alone, TREM1‐LYTAC in the presence of CI‐M6PR blockade, or conjugated TREM1‐LYTAC ( n = 5 independent experiments; one‐way ANOVA followed by Tukey's multiple‐comparison test). (l) Plasma pharmacokinetic profile of free TREM1‐LYTAC after intravenous administration. Fluorescence intensity was fitted to a monoexponential decay model, yielding an apparent half‐life of 1.13 h ( n = 3 mice). (m) Proposed mechanism of TREM1‐LYTAC‐mediated degradation. TREM1‐LYTAC bridges cell‐surface TREM1 to CI‐M6PR, thereby promoting receptor internalization and lysosomal degradation. Data are mean ± SD. Each dot represents an independent experimental replicate or an individual animal. Statistical tests are specified for each quantitative panel above. Exact p values are provided in the Source Data. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐1β ELISA kit , ABclonal , # RK00006.

Techniques: Biomarker Discovery, Binding Assay, Generated, Analysis, Concentration Assay, Drug discovery, Enzyme-linked Immunosorbent Assay, SPR Assay, Confocal, Expressing, Comparison, Functional Assay, Clinical Proteomics, Fluorescence

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Engineering and validation of a BBB‐targeted, pH‐responsive nanodevice for spatiotemporal delivery of TREM1‐LYTAC. (a) Schematic illustration of APTL‐NP fabrication. TREM1‐LYTAC was conjugated to a generation‐4 PAMAM dendrimer through a terephthalaldehyde‐derived acid‐labile linker, followed by surface functionalization with Angiopep‐2 and PS‐PEG‐NHS to enable LRP1‐directed BBB targeting and improve systemic stability. (b) Representative transmission electron microscopy images of PAMAM‐based precursor particles and APTL‐NP (Scale bars, 1 µm). (c) Dynamic light scattering profiles showing the apparent hydrodynamic size distributions of G4 PAMAM and APTL‐NP under the indicated measurement conditions. (d) Time‐dependent changes in hydrodynamic diameter and polydispersity index of APTL‐NP incubated in PBS at pH 7.4 and 37°C. (e) pH‐dependent zeta‐potential profile of APTL‐NP. (f) Nanoparticle tracking analysis showing pH‐dependent particle size and concentration distributions after incubation at pH 7.5, 7.0, or 6.5. (g) pH‐responsive release of TREM1‐binding‐competent TREM1‐LYTAC from APTL‐NP. Released free TREM1‐LYTAC was separated by ultrafiltration and quantified using a TREM1‐binding ELISA with free TREM1‐LYTAC standards ( n = 3 independent experiments). (h) Representative EdU staining images of BMECs, HT22 cells, and NHAs treated with vehicle or APTL‐NP. (i) Quantification of EdU‐positive cells in the indicated cell types( n = 6 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (j) Representative flow‐cytometric plots for apoptosis analysis in BMECs, HT22 cells, and NHAs after treatment with vehicle or APTL‐NP. (k) Quantification of apoptotic cells in the indicated cell types( n = 6 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (l) Cell viability after 24 h or 48 h exposure to the indicated concentrations of APTL‐NP ( n = 3 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (m) Plasma pharmacokinetic profile of APTL‐NP after intravenous administration. The fluorescence signal was fitted to a monoexponential decay model, yielding an apparent half‐life of 8.38 h ( n = 3 mice). (n) Representative photographs and quantification of hemolysis induced by the indicated concentrations of APTL‐NP ( n = 3 independent experiments, one‐way ANOVA followed by Tukey's multiple‐comparison test). (o, p) Plasma C3a and C5a concentrations at the indicated time points after administration of vehicle or APTL‐NP ( n = 6 mice per group, Two‐way ANOVA followed by Tukey's multiple‐comparison test). (q) Representative H&E‐stained sections of the spleen, heart, lung, liver, and kidney from vehicle‐ and APTL‐NP‐treated mice (Scale bar, 100 µm). Data are presented as mean ± SD where applicable. For in vitro assays, n denotes independent experimental replicates; for in vivo analyses, n denotes individual mice. Statistical tests are specified for each quantitative panel above. Exact p values are provided in the Source Data. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: IL‐1β ELISA kit , ABclonal , # RK00006.

Techniques: Biomarker Discovery, Derivative Assay, Stability, Transmission Assay, Electron Microscopy, Dynamic Light Scattering Assay, Incubation, Zeta Potential Analyzer, Analysis, Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Staining, Comparison, Clinical Proteomics, Fluorescence, In Vitro, In Vivo

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Key resources table.

Article Snippet: IL‐1β ELISA kit , ABclonal , # RK00006.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software