human nrg1 alpha  (Novus Biologicals)

Journal: Molecular Cancer Therapeutics

Article Title: AMT-562, a Novel HER3-targeting Antibody–Drug Conjugate, Demonstrates a Potential to Broaden Therapeutic Opportunities for HER3-expressing Tumors

doi: 10.1158/1535-7163.MCT-23-0198

Figure Lengend Snippet: Generation and characterization of Ab562, a novel HER3-targeting antibody. A, Ab562 binds to an epitope located in ECD1 determined by non–cross-reactive mouse HER3 domain swaps. Binding sites of other HER3-targeting mAbs: MM-121 or lumretuzumab (domain I) prevents NRG1 binding, LJM716 binds an epitope created by ECD domains II and IV in the HER3 closed conformation and patritumab binds the extracellular domain II. Modified from Kinisha Gala; Sarat Chandarlapaty. Molecular Pathways: HER3 Targeted Therapy. Clin Cancer Res 20, 1410–1416 (2014). B, Ab562 binding to HER3 in the absence (top) or presence (bottom) of NRG1 measured by BLI analysis. C, Target overexpression (colocalization of red mAb staining and green GFP-tagged target in 293T). Negative staining on the untransfected cells (blue only) is indicated by two red arrows. Scale bars, 10 μm. D, Ab562 caused less cell surface HER3 expression decrease in PC9 measured by flow cytometry. Change of HER3-positive cell percentage is tabulated on the right. Data shown are mean of triplicate measurements and error bars are SEM. Unpaired two-sided t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. E, Effect of Ab562 on HER3 signaling. Western blot analysis for the phosphorylation of HER3, AKT, ERK1/2. Cells were incubated with 30 nmol/L patritumab or Ab562 with or without 1 ng/mL neuregulin-1 (NRG-1) for 1 hour. Western blot results from two ADCs, AMT-562 and P-GGFG-DXd, were also shown here (described in the section “ In vivo functional validation of P-GGFG-DXd/U3-1402 and comparison with AMT-562”). F, In vitro proliferation experiments of Ab562 and patritumab. Cancer cell lines with HER3 and NRG1 expression data (labeled) treated with 100 μg/mL anti-HER3 antibodies for 5 days with cell viability determined by CTG assay. Cell proliferation values are relative to untreated cells and represent average of three replicates ± SEM. Unpaired two-sided t test. *, P < 0.05; **, P < 0.01. G, In vivo efficacy of Ab562 (blue) and patritumab in a breast cancer cell line xenograft model. High HER3 expression in BT-474 was shown as flow cytometry data on the left. Mice were intraperitoneally administered with antibodies (20 mg/kg) and on day 0 (tumor size reached an average of 150–200 mm 3 ) and subsequent dates indicated by red arrows. Each value represents the mean and SEM ( n = 5). Unpaired two-sided t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Binding measurement was performed in the absence or presence of NRG1 (13499-H08H, SinoBiological).

Techniques: Binding Assay, Modification, Over Expression, Staining, Negative Staining, Expressing, Flow Cytometry, Western Blot, Incubation, In Vivo, Functional Assay, Comparison, In Vitro, Labeling, CTG Assay

Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) CD11c + cells are increased in the lungs of atopic mice. Flow cytometry of lung cell suspension showing frequency (left) and cell number (right) at day 3 post inoculation (PI) high dose SeV ( B ) Adoptive transfer of CD11c + cells isolated from NA or atopic mice into naïve mice 24 h before inoculation with high dose SeV delays but does not prevent mortality; n=4 per group. ( C ) Transcriptomic (RNAseq) comparison between FACS isolated lung CD11c + cells from atopic and NA mice identifies several disparately expressed gene products, including Nrg1 (n=4 per group). Selected gene products shown. ( D ) NRG1 is markedly increased in atopic mouse lung (“tissue”), BAL, and supernatant from 1 x10 6 atopic lung CD11c + cells (“CD11c sup”) cultured for 24 h. ( E ) The scRNA (10x Platform) tSNE coordinates show CD14 + monocytes and dendritic cell (“DC”) sub-populations expressing NRG1 in peripheral blood cells of healthy human donors.*p<0.05, **p<0.01, ****p<0.0001

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: Flow Cytometry, Suspension, Adoptive Transfer Assay, Isolation, Comparison, Cell Culture, Expressing

Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) NRG1(1ng to 1000ng) i.n. (in 30µL) given daily to naïve mice for 5d before inoculation with high dose SeV reduces viral mortality; n=4 per group (1ng, 10ng 1000ng & PBS), n=8 per group (100 ng and 500 ng). ( B ) Ratio of EBD in the BAL to that in the lung shows reduced EBD in NRG1 treated mice on day 8 PI SeV. n≥8 per group, median ± IQR shown, Mann-Whitney U test.

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: MANN-WHITNEY

Journal: bioRxiv

Article Title: Neuregulin-1 protects against respiratory viral induced mortality

doi: 10.1101/2023.05.10.540232

Figure Lengend Snippet: ( A ) Adding NRG1 to hBEC inoculated with rgRSV (left) and mTEC inoculated with GFP-SeV (right) reduces spread of infection. Representative images shown. ( B ) Quantification of (A) for rgRSV and hBEC and ( C ) for GFP-SeV and mTEC. GFP positive cells quantified by ImageJ. Representative images from ≥3 separate experiments. *p<0.05, **p<0.01. ( D ) Transcriptomic analysis of hBEC cultures treated with NRG1 (100 ng) on the basolateral side of the Transwell for 5 days and inoculated with RSV (4000 pfu). RNA was isolated 48 h PI RSV and qRT-PCR performed using a custom Prime PCR array plate: (i) Transcripts in which NRG1 treatment reduced gene expression from that seen in RSV infected cells. (ii) Transcripts with low level expression that show small but significant change in expression relative to naïve control with NRG1 alone or genes significantly increased with RSV but whose expression levels were not affected by NRG1. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, n=3.

Article Snippet: Cell culture basal media was supplemented with recombinant human NRG1 alpha (cat#: NBP2-35093, Novus Bio) for hBEC or with mouse NRG1 for mTEC on day five and three before and at the time of infection with 4000 pfu of rgRSV or SeV-GFP.

Techniques: Infection, Isolation, Quantitative RT-PCR, Expressing, Control

Journal: Science signaling

Article Title: Kinetics of receptor tyrosine kinase activation define ERK signaling dynamics *

doi: 10.1126/scisignal.aaz5267

Figure Lengend Snippet: (A) Endogenous ErbB4 in MCF7 cells was stimulated using saturating levels of NRG1β (25 nM/ 200 ng/ml) or NRG1α (1 μM/ 7 μg/ml). Activating phosphorylation of ErbB4 and ERK1/2, as well as total ErbB4 and ERK levels, were monitored by immunoblotting total cell lysates. Grb2 was used as a loading control. Blots are representative of at least three independent experiments. Statistical significance of differences was assessed using the Bonferroni-Dunn method (alpha = 0.05), with datasets for each point analyzed individually without assuming consistent SD.

Article Snippet: Recombinant human NRG1α (Cat. #296-HR) and NRG1β (Cat. #396-HB/CF) were obtained from R&D Systems.

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Science signaling

Article Title: Kinetics of receptor tyrosine kinase activation define ERK signaling dynamics *

doi: 10.1126/scisignal.aaz5267

Figure Lengend Snippet: (A) Endogenous ErbB4 in MCF7 cells was stimulated using saturating levels of NRG1β (25 nM/ 200 ng/ml) or NRG1α (1 μM/ 7 μg/ml). Activating phosphorylation of ErbB4 and ERK1/2, as well as total ErbB4 and ERK levels, were monitored by immunoblotting total cell lysates. Grb2 was used as a loading control. Blots are representative of at least three independent experiments. Statistical significance of differences was assessed using the Bonferroni-Dunn method (alpha = 0.05), with datasets for each point analyzed individually without assuming consistent SD.

Article Snippet: Recombinant human NRG1α (Cat. #296-HR) and NRG1β (Cat. #396-HB/CF) were obtained from R&D Systems.

Techniques: Phospho-proteomics, Western Blot, Control