Journal: Molecular cancer therapeutics

Article Title: Development of antibody-drug conjugates targeting L1CAM to treat metastatic cancer

doi: 10.1158/1535-7163.MCT-25-1184

Figure Lengend Snippet: L1CAM as a prognostic marker and a therapeutic target in various cancer types. A , L1CAM transcript levels in primary and metastatic tumors from different tissues. Bar graph, mean ± S.E.M. Sample sizes are indicated in the figure. Data were sourced from TNMplot. Scheme created with BioRender. Tumor L1CAM expression is likely to be underestimated in stroma-dense tumors (e.g. Ovarian, Pancreatic). B , The overall survival rate for various cancer types with respect to L1CAM transcript levels. Overall survival, individual data points in grey; median survival in black and red. Sample sizes are indicated in the figure. Data were sourced from KM-plotter which integrates datasets from GEO, EGA, and TCGA. C , L1CAM IHC staining of LUAD primary tumor and metastases from patient tissue sections. Scale bar, 50 μm. D , Clinical profiles of LUAD PDX cohort ranked by L1CAM H-score. Each column represents a single patient from whom the xenograft was derived. Sample sizes are indicated in the figure. E , L1CAM IHC staining of LUAD PDX tissue section, followed by immunofluorescence imaging of L1CAM staining and UMAP analysis of L1CAM transcript levels from scRNA-seq data (4,017 cells) from PDX tumoroids. Scale bar, 30 μm (IHC); 10 μm (IF). Selected PDX line is indicated by the black arrow from panel D. F , L1CAM mRNA levels from a bulk RNA-seq in various LUAD cell lines and their KRAS mutation status. Each column represents a single cell line. Sample sizes are indicated in the figure. A.U., arbitrary unit. G , Western immunoblot analysis of L1CAM in selected LUAD cell lines from panel F , as indicated by the colored arrows. Statistical significance was assessed using a two-tailed Mann-Whitney test ( A,B ). ns, not significant; * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: Antibodies used include mouse L1CAM (Miltenyi Biotec Cat# 130-115-812, AB_2727206), human L1CAM (Santa Cruz Biotechnology Cat# sc-53386, RRID: AB_628937), Sox2 (Invitrogen Cat# 14-9811-82, RRID: AB_891383), GFP (Aves Labs Cat# GFP-1010, RRID: AB_2307313), Cleaved Caspase-3 (Cell Signaling Technology Cat# 9661, RRID: AB_2341188), and human cytokeratin (Dako Cat# M3515, RRID: AB_2132885).

Techniques: Marker, Expressing, Immunohistochemistry, Derivative Assay, Immunofluorescence, Imaging, Staining, RNA Sequencing, Mutagenesis, Single Cell, Western Blot, Two Tailed Test, MANN-WHITNEY

Journal: Molecular cancer therapeutics

Article Title: Development of antibody-drug conjugates targeting L1CAM to treat metastatic cancer

doi: 10.1158/1535-7163.MCT-25-1184

Figure Lengend Snippet: Generation of L1CAM antibodies binding to the extracellular portion. A , Schematic representation of the L1CAM molecule and its extracellular domains as binding sites for the generated antibodies. B , Workflow for hit selection of hybridoma supernatants from mice immunized with L1CAM immunogens. C , Molecular characterization of the L1CAM antibodies derived from the experimental steps outlined in panel B .

Article Snippet: Antibodies used include mouse L1CAM (Miltenyi Biotec Cat# 130-115-812, AB_2727206), human L1CAM (Santa Cruz Biotechnology Cat# sc-53386, RRID: AB_628937), Sox2 (Invitrogen Cat# 14-9811-82, RRID: AB_891383), GFP (Aves Labs Cat# GFP-1010, RRID: AB_2307313), Cleaved Caspase-3 (Cell Signaling Technology Cat# 9661, RRID: AB_2341188), and human cytokeratin (Dako Cat# M3515, RRID: AB_2132885).

Techniques: Binding Assay, Generated, Selection, Derivative Assay

Journal: Molecular cancer therapeutics

Article Title: Development of antibody-drug conjugates targeting L1CAM to treat metastatic cancer

doi: 10.1158/1535-7163.MCT-25-1184

Figure Lengend Snippet: Payload optimization for L1CAM ADCs. Cytotoxicity curves for L1CAM ADCs generated from either the 14A10 (purple) or 143G03 (red) antibody conjugated to PNU-159682 ( A ), MMAE ( B ), ABZ038 ( C ), Duocarmycin ( D ), or α-Amanitin ( E ) using a Val-Cit linker. Cell viability was measured as a function of ADC concentration (0.1 pM to 100 nM) after 96 h incubation with HEK293T cells overexpressing L1CAM (293T-L1CAM) or MCF-7 and MDA-MB-231 human breast cancer cell lines. A mouse IgG1 isotype control antibody conjugated to the corresponding payload (black) and the corresponding free drug (dotted grey) were included as controls.

Article Snippet: Antibodies used include mouse L1CAM (Miltenyi Biotec Cat# 130-115-812, AB_2727206), human L1CAM (Santa Cruz Biotechnology Cat# sc-53386, RRID: AB_628937), Sox2 (Invitrogen Cat# 14-9811-82, RRID: AB_891383), GFP (Aves Labs Cat# GFP-1010, RRID: AB_2307313), Cleaved Caspase-3 (Cell Signaling Technology Cat# 9661, RRID: AB_2341188), and human cytokeratin (Dako Cat# M3515, RRID: AB_2132885).

Techniques: Generated, Concentration Assay, Incubation, Control

Journal: Molecular cancer therapeutics

Article Title: Development of antibody-drug conjugates targeting L1CAM to treat metastatic cancer

doi: 10.1158/1535-7163.MCT-25-1184

Figure Lengend Snippet: Linker optimization for L1CAM ADCs. Schematic representations of the ADC with cleavable Glu-Val-Cit ( A ) or non-cleavable tri-glycine ( B ) linkers and the PNU-159682 payload ( 50 ). Cytotoxicity curves for 143G03 (red) and 14A10 (blue) ADCs conjugated to PNU-159682 with either a cleavable ( C ) or non-cleavable ( D ) linker in 293T-L1CAM, MCF-7, and MDA-MB-231 cell lines. An ADC generated from a mouse IgG1 isotype control antibody (IgG1-ADC; black) and free PNU-159682 (dotted grey) were included as controls.

Article Snippet: Antibodies used include mouse L1CAM (Miltenyi Biotec Cat# 130-115-812, AB_2727206), human L1CAM (Santa Cruz Biotechnology Cat# sc-53386, RRID: AB_628937), Sox2 (Invitrogen Cat# 14-9811-82, RRID: AB_891383), GFP (Aves Labs Cat# GFP-1010, RRID: AB_2307313), Cleaved Caspase-3 (Cell Signaling Technology Cat# 9661, RRID: AB_2341188), and human cytokeratin (Dako Cat# M3515, RRID: AB_2132885).

Techniques: Generated, Control

Journal: Molecular cancer therapeutics

Article Title: Development of antibody-drug conjugates targeting L1CAM to treat metastatic cancer

doi: 10.1158/1535-7163.MCT-25-1184

Figure Lengend Snippet: In vivo efficacy of L1CAM ADCs against a triple negative breast cancer model. Athymic mice were transplanted subcutaneously ( A, B ) or via tail vein injection to establish orthotopic lung metastasis ( C, D ) with MDA-MB-231 LM2 breast cancer cells. Once tumors had grown, mice were administered 4 weekly doses (shown in red arrows) of ADCs generated with the 143G03 or 14A10 ADC conjugated to PNU-159682 with a cleavable (C) linker, (0.3 mg/kg) or non-cleavable (NC) linker (1 mg/kg), and IgG1-ADC or vehicle control. Tumor volume ( A ), Weekly lung metastasis ( C ) and overall survival ( B, D ) were monitored. n = 6 mice per condition. Statistical significance was assessed using the Kolmogorov-Smirnov test ( A, C ) or log-rank (Mantel-Cox) test ( B, D ). ns, not significant; * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: Antibodies used include mouse L1CAM (Miltenyi Biotec Cat# 130-115-812, AB_2727206), human L1CAM (Santa Cruz Biotechnology Cat# sc-53386, RRID: AB_628937), Sox2 (Invitrogen Cat# 14-9811-82, RRID: AB_891383), GFP (Aves Labs Cat# GFP-1010, RRID: AB_2307313), Cleaved Caspase-3 (Cell Signaling Technology Cat# 9661, RRID: AB_2341188), and human cytokeratin (Dako Cat# M3515, RRID: AB_2132885).

Techniques: In Vivo, Injection, Generated, Control

Journal: Molecular cancer therapeutics

Article Title: Development of antibody-drug conjugates targeting L1CAM to treat metastatic cancer

doi: 10.1158/1535-7163.MCT-25-1184

Figure Lengend Snippet: In vitro efficacy of L1CAM ADC across various cancer types. Cytotoxicity curves for 13F04 ADC (red) conjugated to PNU-159682 with a non-cleavable linker in the following cell lines: 293T-L1CAM ( A ), MCF-7 ( B ), MDA-MB-231 ( C ), H524 ( D ), H2030 ( E ), H23 parental ( F ), and H23 with L1CAM knockout ( G ). Efficacy was also evaluated in PDX-derived Ru631 tumoroids with either shControl ( H ) or shL1CAM knockdown ( I ). An ADC generated from a mouse IgG1 isotype control antibody (IgG1-ADC; black) was used as a negative control.

Article Snippet: Antibodies used include mouse L1CAM (Miltenyi Biotec Cat# 130-115-812, AB_2727206), human L1CAM (Santa Cruz Biotechnology Cat# sc-53386, RRID: AB_628937), Sox2 (Invitrogen Cat# 14-9811-82, RRID: AB_891383), GFP (Aves Labs Cat# GFP-1010, RRID: AB_2307313), Cleaved Caspase-3 (Cell Signaling Technology Cat# 9661, RRID: AB_2341188), and human cytokeratin (Dako Cat# M3515, RRID: AB_2132885).

Techniques: In Vitro, Knock-Out, Derivative Assay, Knockdown, Generated, Control, Negative Control

Journal: Molecular cancer therapeutics

Article Title: Development of antibody-drug conjugates targeting L1CAM to treat metastatic cancer

doi: 10.1158/1535-7163.MCT-25-1184

Figure Lengend Snippet: In vivo efficacy of L1CAM ADC against triple negative breast cancer and lung adenocarcinoma (LUAD) metastasis. A,B , Single-cell suspensions of MDA-MB-231 LM2 cells transduced with luciferase were injected into NSG mice via the tail vein. After 1 week of metastatic engraftment and outgrowth in the lungs, the mice received control ADC (1 mg/kg) or 13F04 ADC (1 mg/kg) by intraperitoneal injection once weekly for four weeks at the indicated times ( red arrows ). Tumor burden was monitored by BLI weekly. A , Metastatic burden of mice inoculated with MDA-MB-231 cells and treated with control ADC or 13F04 anti-L1CAM ADC. n = 5 mice per group. *** P = 0.0006. B , Overall survival plots of mice in the experiment of panel ( A ) ** P = 0.0035. C , NSG mice were inoculated via the tail vein with GFP-expressing Ru631 LUAD PDX cells. One week later, mice were treated with a single dose of 13F04 ADC or IgG control ADC, and lungs were harvested and analyzed by IF staining for GFP (cancer cells) and L1CAM in one week after ADC treatment. White arrows, cytoplasmic L1CAM staining upon anti-L1CAM ADC treatment. Scale bar, 10 μm. D , Quantification of L1CAM − or L1CAM + fraction of Ru631 PDX treated with ADC in the experiment of panel ( C ). n = 3,195 cells (Control); 2,110 cells (13F04). ** P = 0.0079. E , H&E staining of lung sections before or one week after a single dose of ADC treatment, administered four weeks after inoculation of Ru631 PDX cells into NSG mice via the tail vein. Magnified regions are shown ( red boxes ). Scale bar, 200 μm. F , Bar graph showing the percent area of metastatic lesions per lung in the experiments of panel ( E ). n = 3 per experimental condition. * P = 0.0239. G , IF staining for human pan-cytokeratin (cancer cells) and cleaved caspase 3 in lung metastatic colonies one week after a single dose of ADC treatment, administered four weeks post-inoculation of Ru631 PDX cancer cells into NSG mice via the tail vein. Scale bar, 50 μm. H , Fraction of cleaved caspase 3 + (apoptotic) cells detected by IF in the experiment of panel ( G ). n = 3,257 cells (Control); 788 cells (13F04). **** P < 0.0001. I , IF staining for GFP (cancer cells) and SOX2 one week after a single dose of ADC treatment, administered one week after inoculation of Ru631 PDX cancer cells into NSG mice via the tail vein. Scale bar, 10 μm. J , Percentage of SOX2 + cancer cells detected by IF in the experiment of panel ( I ). n = 5 per experimental condition. * P = 0.0317. K-N , Therapeutic assessment of LUAD metastasis treated with L1CAM or IgG ADCs. Single-cell suspensions from LUAD PDX tumoroids transduced with luciferase were injected into NSG mice via the tail vein. After 4 to 5.5 weeks of metastatic engraftment and outgrowth in the lungs, the mice received control ADC (1 mg/kg) or 13F04 ADC (1 mg/kg) by intraperitoneal injection once weekly for four weeks at the indicated times ( red arrows ). Tumor burden was monitored by BLI weekly. K , Metastatic burden of mice inoculated with Ru323 PDX tumoroid cells and treated with control ADC or 13F04 anti-L1CAM ADC. n = 5 mice per group. **** P < 0.0001. L , Overall survival plots of mice in the experiment of panel ( K ) ** P = 0.0018. M , Metastatic burden of mice inoculated with Ru631 PDX tumoroid cells and treated with control ADC or 13F04 anti-L1CAM ADC. Control, n = 14 mice; 13F04, n = 15 mice. **** P < 0.0001. N , Overall survival plots of mice in the experiment of panel ( M ). **** P < 0.0001. The bar graph indicates mean ± S.E.M. ( D,F,J ). Data are shown as a box (median ± 25-75%) and whisker (maximum to minimum values) plot ( H ). Statistical significance was assessed using the two-tailed Mann-Whitney test ( D,H,J ), two-tailed t test after passing the Shapiro-Wilk normality test ( F ), Kolmogorov-Smirnov test ( A,K,M ) or log-rank (Mantel-Cox) test ( B,L,N ).

Article Snippet: Antibodies used include mouse L1CAM (Miltenyi Biotec Cat# 130-115-812, AB_2727206), human L1CAM (Santa Cruz Biotechnology Cat# sc-53386, RRID: AB_628937), Sox2 (Invitrogen Cat# 14-9811-82, RRID: AB_891383), GFP (Aves Labs Cat# GFP-1010, RRID: AB_2307313), Cleaved Caspase-3 (Cell Signaling Technology Cat# 9661, RRID: AB_2341188), and human cytokeratin (Dako Cat# M3515, RRID: AB_2132885).

Techniques: In Vivo, Single Cell, Transduction, Luciferase, Injection, Control, Expressing, Staining, Whisker Assay, Two Tailed Test, MANN-WHITNEY

Journal: Experimental Biology and Medicine

Article Title: Curcumin-enhanced elvitegravir therapy mitigates neuroinflammation and cognitive deficits in EcoHIV mice

doi: 10.3389/ebm.2025.10758

Figure Lengend Snippet: Brain levels of 8-OHdG (A,B) and glutamate (C,D) were measured in EcoHIV-infected mice treated with EVG, CUR, or their combination (EVG + CUR) via IN or IP routes. (E–I) Western blot analysis was conducted to assess the expression of neural protein markers NeuN, TMEM119, synaptophysin, L1CAM, and GFAP in the mice brains. Data are expressed as mean ± SEM (n = 4 per group). Statistical analysis was performed by one-way ANOVA with Tukey’s post hoc test. *, **, and *** represents p ≤ 0.05, p ≤ 0.01, and p ≤ 0.001 respectively.

Article Snippet: Primary antibody incubation was carried out overnight at 4 °C using the following antibodies: NeuN rabbit polyclonal antibody (1:1000, Proteintech, Cat# 26975-1-AP), synaptophysin mouse monoclonal antibody (1:20,000, Proteintech, Cat# 67864-1-Ig), GFAP rabbit polyclonal antibody (1:1000), L1CAM rabbit polyclonal antibody (Proteintech, Cat# 20659-1-AP), and β-actin mouse monoclonal antibody (1:20,000, Proteintech, Cat# 66009-1-Ig) as an internal loading control.

Techniques: Infection, Western Blot, Expressing