elabscience e el h0114  (Elabscience Biotechnology)

Journal: Nature Communications

Article Title: A yeast surface display platform for characterizing CAR T cell responses to cancer antigens

doi: 10.1038/s41467-025-65236-7

Figure Lengend Snippet: a Heterologous G protein-coupled receptors (GPCR) as sensory modules for controlling CD19 display with cognate agonists: fold change CD19 levels ( top row ) and absolute CD19 histograms ( lower row ). All strains employ P FUS1 and vary between GPCRs and G ɑ -subunits. b Comparison of CD19 levels across GPCRs. c . CD19 SCASA yeast design; CD19 expression is controllable by GPCR-dependent ligands ( sensory module ). System regulation depends on signaling through an engineered pheromone response pathway (PRP) via G ɑβγ -protein, choice of promoter, and expression boost effects ( processing module ). CD19 output is a fusion protein composed of HA- and myc-tags, PAS40-linker, (G 4 S) 3 -linker, and CD19.1 ECD, fused to Aga2 ( effector module ). Strains were optimized by; G ɑ -subunit, GPCR, and Aga1 overexpression, as well as gene knock-outs; ste2Δ0, ste3Δ0, gpa1Δ0, sst2Δ0, bar1Δ0, far1Δ0, aga2Δ0 . Created in BioRender. Deichmann, M. (2025): https://BioRender.com/dtuelpw . d Relative yEGFP levels of promoters with ɑ-factor stimulation including PRP boost effects, sorted; low to high (5 h. post-induction). e Fold change promoter induction (yEGFP), excluding PRP boost effects (SSC-normalization), sorted; low to high. f Approximated PRP boost of yEGFP expression from quantified PRP activation ( black ) across all strains and designs ( grey ) ( n = 27). g CD19 histograms of SCASA yeast with different promoters without GPCR stimulation ( green ), a P PGK1 -Empty control lacking CD19 ( red ), and NALM6 ( blue ). h Fold change CD19 of SCASA yeast during GPCR stimulation with ɑ-factor (20 h. post-induction). i Approximated PRP boost of CD19 display from quantified PRP activation ( black ), across all strains and designs ( grey ) ( n = 15). j Comparison of CD19 levels during GPCR stimulation of SCASA yeast, relative to lowest detected CD19 level ( top row ), and CD19 histograms ( lower row ). Unless otherwise noted, data is means of median fluorescence intensities (mMFI) for biological replicates ( n = 3) and standard deviations hereof. Histograms are representative replicates normalized to the mode. Statistical tests: One- and Two-way ANOVA with multiple comparisons statistical tests. Significance levels: * p ≤ 0.05, ** p ≤ 0.001. Not all pairwise comparisons are shown. All statistics and extended analyses in: Supplementary Figs. – and Supplementary Data – . Source data provided as a Source Data file.

Article Snippet: The human CD19+ NALM6 B cell precursor leukemia cell line (DSMZ, no.: ACC 128) was included as a benchmark positive control.

Techniques: Comparison, Expressing, Over Expression, Activation Assay, Control, Fluorescence

Journal: Nature Communications

Article Title: A yeast surface display platform for characterizing CAR T cell responses to cancer antigens

doi: 10.1038/s41467-025-65236-7

Figure Lengend Snippet: a Illustration of chimeric antigen receptor (CAR) Jurkat cell co-culture with SCASA yeast cells. Jurkat cells that express anti-CD19 CARs ( CAR + ) bind to CD19+ SCASA yeast cells, analogously to as for the CD19+ NALM6 human cancer cell line. Here, Jurkat cells contain a NFAT-Luc reporter system, which upon activation expresses luciferase. SCASA yeast cells can be stimulated through their G protein-coupled receptors (GPCR) to induce differential CD19 antigen density (e.g. Ste2 with ligand ɑ-factor). Created in BioRender. Deichmann, M. (2025): https://BioRender.com/urlsdg9 . b Relative CAR activation in co-cultures of 100% CAR+ Jurkat NFAT-Luc cells with SCASA yeast designs; P FUS1 -CD19, P MFA2 -CD19, and P TDH3 -CD19 ( green lines ) at a target-to-effector (T/E) cell ratio of 0.2x yeast cell per Jurkat cell for 18 h., with increasing SCASA yeast GPCR stimulation for CD19 display control (ɑ-factor). Negative control: P PGK1 -Empty lacking the CD19 CDS in the display construct ( red line ). Positive benchmark control: NALM6 ( blue line ). One-way ANOVA with Dunnett’s multiple comparisons statistical test of NFAT-activation compared to 0 µM is shown for each strain. c Relative CAR activation (log 10 -scale) in co-cultures of Jurkat NFAT-Luc cells with unstimulated SCASA yeast designs; P MFA2 -CD19, P PGK1 -CD19, and P TDH3 -CD19, as well as a negative control P PGK1 -Empty, and NALM6 as benchmark control. The percentage of Jurkat NFAT-Luc cells expressing CARs was graded from 0–100% CAR+ . Two-way ANOVAs are shown for a comparison of background activation at 0% CAR+ (Tukey’s multiple comparisons test) ( box ), and significant activation within the 0–100% CAR+ range for each SCASA yeast strain and NALM6 relative to 0% CAR+ (Dunnett’s multiple comparisons test). Data is based on means of three biological replicates ( n = 3) and standard deviations hereof. Significance levels: ns : not significant, * p ≤ 0.05, ** p ≤ 0.001. Not all pairwise comparisons are shown. All statistics and extended analyses in: Supplementary Fig. and Supplementary Data . Source data provided as a Source Data file.

Article Snippet: The human CD19+ NALM6 B cell precursor leukemia cell line (DSMZ, no.: ACC 128) was included as a benchmark positive control.

Techniques: Co-Culture Assay, Activation Assay, Luciferase, Control, Negative Control, Construct, Expressing, Comparison

Journal: Nature Communications

Article Title: A yeast surface display platform for characterizing CAR T cell responses to cancer antigens

doi: 10.1038/s41467-025-65236-7

Figure Lengend Snippet: a Chimeric antigen receptor (CAR) T cell designs FMC63-CD28-CD28-CD3ζ (axicabtagene ciloleucel/brexucabtagene autoleucel) (‘CD28 CAR’) and FMC63-CD8ɑ−4-1BB-CD3ζ (tisagenlecleucel) (‘4–1BB CAR’), expressed in a triple-parameter-reporter (TPR) Jurkat T-cell line that couple T-cell activation transcription factors to fluorescent outputs; NF-κB-CFP, NFAT-eGFP, and AP-1-mCherry. CAR TPR Jurkats were co-cultivated for 24 h. with six CD19 SCASA yeast strains with different processing modules and hence different baseline CD19 levels. Antigen-density modulation was examined by G protein-coupled receptor (GPCR) stimulation of individual strains (ɑ-factor) at a 1.0x target-to-effector (T/E) cell ratio. The effects of increasing target cell numbers were examined by modulating the T/E-ratio of SCASA yeast strains with fixed antigen densities and NALM6 from x0.25 to x8.0 relative to a fixed amount of CAR T cells. Created in BioRender. Deichmann, M. (2025): https://BioRender.com/65vslu2 . b Relative response intensity of NF-κB-CFP ( blue ), NFAT-eGFP ( green ), and AP-1-mCherry ( red ) for the CD28 CAR ( light ) and 4-1BB CAR ( dark ) across all examined conditions, normalized to 4-1BB monocultures (log 2 -scale). ɑCD3/ɑCD28 dynabeads (1.0x) were employed as a positive control for reporter genes. c Comparison between yeast and microbeads for CAR activation. CAR activation relative to the amount of CD19 molecules per target (log 10 -scale) with comparison between equal numbers of SCASA yeast cells ( circle ) and CD19-coated microbeads ( triangle ) (1.5x T/E-ratio), measured by relative response intensity (log 2 -scale). Microbeads had a 5.5 µm diameter, were streptavidin coated, and loaded with biotinylated CD19 (Supplementary Fig. ). A comparison to CD19-coated microtiter plates was also done, however, the planar and continuous surface is physically distinct from yeast and microbeads, as well as with unknown antigen densities (Supplementary Fig. ). Data represents means of median fluorescence intensities (mMFI) for three biological replicates ( n = 3) and standard deviations hereof. All statistics and extended analyses in: Supplementary Figs. – and Supplementary Data – . Source data are provided as a Source Data file.

Article Snippet: The human CD19+ NALM6 B cell precursor leukemia cell line (DSMZ, no.: ACC 128) was included as a benchmark positive control.

Techniques: Activation Assay, Positive Control, Comparison, Fluorescence

Journal: Nature Communications

Article Title: A yeast surface display platform for characterizing CAR T cell responses to cancer antigens

doi: 10.1038/s41467-025-65236-7

Figure Lengend Snippet: a CD69 expression in non-engineered control (CTRL) T cells (alive, CD3+ ) ( upper ) and chimeric antigen receptor (CAR) T cells (alive, CD3+ , CAR+ ) ( lower ) after co-cultivation with NALM6 ( blue ), SCASA yeast P PGK1 -CD19 ( green ), and negative control yeast P PGK1 -Empty lacking CD19 ( red ), at different target-to-effector (T/E) cell ratios (0.2x, 1.0x, 5.0x) for 20 h. An overlay for 5.0x co-cultivations is shown ( far right ), including unstimulated T cell monocultures ( black line ). b CD69 expression intensity of co-cultivation populations: CAR+ ( circle ) and CAR- ( square ) T cells from the CAR-engineered population, CTRL T cells ( triangle ), and baseline CD69 of monocultures ( dotted lines ). Normalization: CTRL T cell monoculture. c Percentage of T cell populations expressing CD69 (%CD69+ ) at any intensity for co-cultivations, and baseline %CD69+ of monocultures ( dotted lines ). d Target cell CD19 levels after co-cultivations at 5.0x for NALM6 and P PGK1 -CD19 yeast with CAR T cells ( light ) and CTRL T cells ( dark ). e CD19 levels relative to maximum of NALM6 and P PGK1 -CD19 yeast individually after co-cultivations with CTRL T cells ( triangle ) and CAR T cells ( circle ). f Fold target cell number variation per alive CD3+ T cell between CTRL and CAR T cell cultures for NALM6, P PGK1 -CD19, and P PGK1 -Empty, disregarding increased or diminished levels of cells (reciprocal value of fold changes<1). g CAR expression intensity of CAR+ population relative to CAR- population (background), normalized to maximum. h Percentage of CAR+ T cells per CD3+ T cell detected after co-cultivations. i Comparison of general performance of P PGK1 -CD19 SCASA yeast and NALM6 cancer cells, quantified as percentage of most extreme behavior observed (% of max), averaging across T/E ratios. Summary of all other plots ( a – h ). Data represents means of cell counts or median fluorescence intensities (mMFI) for three biological replicates ( n = 3) and standard deviations hereof. Histograms are representative replicates and normalized to mode (CD69) or unit area (CD19). Selected comparisons from two-way ANOVAs with Tukey’s multiple comparisons tests are shown. Significance levels: * p ≤ 0.05, ** p ≤ 0.001. Statistics and extended analyses in: Supplementary Fig. – and Supplementary Data . Source data provided as a Source Data file.

Article Snippet: The human CD19+ NALM6 B cell precursor leukemia cell line (DSMZ, no.: ACC 128) was included as a benchmark positive control.

Techniques: Expressing, Control, Negative Control, Comparison, Fluorescence

Journal: Scientific Reports

Article Title: The diagnostic value of LncRNA NEAT1 targeting miR-129-5p in pancreatic cancer patients

doi: 10.1038/s41598-025-12963-y

Figure Lengend Snippet: The box plots of serum NEAT1 and miR-129-5p fold changes, BCL2, and TGF-β1 protein levels in control subjects and PC cases. BCL2: B-cell lymphoma-2; NEAT1: Nuclear paraspeckle assembly transcript 1; TGF-β1: Transforming growth factor beta-1; PC: Pancreatic cancer. *: P < 0.05 indicating statistical significance.

Article Snippet: Serum BCL2 and TGF-β1 were investigated using a sandwich enzyme linked immunosorbent assay (ELISA) employing the Human BCL2 (Catalog no: E-EL-H0114) and TGF-β1 (Catalog no: E-EL-H0110) ELISA kits from Elabscience ® , Texas, USA, according to the manufacturer’s directions.

Techniques: Control

Journal: Scientific Reports

Article Title: The diagnostic value of LncRNA NEAT1 targeting miR-129-5p in pancreatic cancer patients

doi: 10.1038/s41598-025-12963-y

Figure Lengend Snippet: ROC curve for serum levels of NEAT1, miR-129-5p, BCL2, TGF-β1, CA19-9 and CEA as indicators for PC diagnosis. BCL2: B-cell lymphoma-2; CEA: Carcinoembryonic antigen; NEAT1: Nuclear paraspeckle assembly transcript 1; CA19-9: cancer antigen 19−9; TGF-β1: Transforming growth factor beta-1.

Article Snippet: Serum BCL2 and TGF-β1 were investigated using a sandwich enzyme linked immunosorbent assay (ELISA) employing the Human BCL2 (Catalog no: E-EL-H0114) and TGF-β1 (Catalog no: E-EL-H0110) ELISA kits from Elabscience ® , Texas, USA, according to the manufacturer’s directions.

Techniques: Biomarker Discovery