Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 increased after conventional chemotherapy regimen Vrd or Krd. A and B. GSE276561 and GSE47552 datasets revealed ALDH1A1 in MM was lower than healthy control; These GEO data were analyzed by on line tool GEO2R using Benjamini & Hochberg (False discovery rate) method; Each y axis label in the graph represents the expression measurement extracted from the TPM normalized expression counts (for RNA-seq), or the Value column of the original submitter-supplied Sample record (for microarrays). All the y axis label represents normalized mRNA expression level. C. mRNA levels of ALDH1A1 in NDMM samples are significantly lower than healthy control, but remarkably higher in RRMM samples than NDMM patients which are collected in our hospital; Relative ALDH1A1 levels is relative to GAPDH. D. mRNA levels of CD38 in NDMM patients are significantly higher than healthy control which are collected in our hospital; Relative CD38 levels is relative to GAPDH. E. ALDH1A1 mRNA increased after Vrd or Krd treatment; Relative ALDH1A1 levels is relative to GAPDH. F . western blot confirmed ALDH1A1 protein increased after Vrd or Krd treatment. **, p < 0.01, ***, p < 0.001. C-E, Data was analyzed by Mann-Whitnay test HC: healthy control; Vrd: Bortezomib + Lenalidomide + Dexamethasone; KRd: Carfilzomib + Lenalidomide + Dexamethasone; MM: multiple myeloma; NDMM: newly diagnosed multiple myeloma; SMM: smoldering multiple myeloma; MGUS: monoclonal gammopathy of undetermined significance

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: Control, Expressing, RNA Sequencing, Western Blot

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 down-regulated in the nrRRMM patients received Dara-Rd treatment. A and B. rRRMM patients had higher ALDH1A expression compared with nrRRMM after Dara-Rd treatment analyzed by RT-qPCR and western blot; C and D . the CD38 expression (mRNA and protein) decreased in Dara-treated patients in nrRRMM patients compared with rRRMM patients; E . the mRNA expression of ALDH1A1 after Dara-based treatment was positively correlated with the CD38 mRNA expression. **, p < 0.01 A-D, data was analyzed by student t test. E, data was analyzed by simple linear regression nrRRMM: non-responder relapsed/refractory multiple myeloma; rRRMM: responder relapsed/refractory multiple myeloma; MFI: median fluorescence intensity; Dara-Rd: daratumumab + lenalidomide + dexamethasone; Dara: daratumumab

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Fluorescence

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 directly regulates CD38 expression in H929 cells. A . the KD efficiency of different sh-ALDH1A1 sequences; B . Knock down ALDH1A1 decreased CD38 genes expression; C. Re-introducing ALDH1A1-OE plasmid restored downregulation of ALDH1A1 mRNA levels in ALDH1A1-KD cells; D . Re-introducing ALDH1A1 into ALDH1A1-KD cells significantly upregulate the expression of CD38; E . cell-surface expression of CD38 decreased in ALDH1A1-KD cells but restored by re-introducing ALDH1A1-OE plasmid determined by flow cytometry analysis. **, p < 0.01, ***, p < 0.001 Student t test was performed in all datasets AlDH1A1-KD: ALDH1A1 knockdown; Ctrl: control; shALDH1A1: short hairpin RNA for ALDH1A1; KD-OE: over expression of ALDH1A1 in KD cells

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: Expressing, Knockdown, Plasmid Preparation, Flow Cytometry, Control, shRNA, Over Expression

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 increased CD38 through activation of RA/RAR via RA production. A . Volcano plots created by differential mRNA (fold change (FC) > 2 or < 0.5, and p < 0.05); B. KEGG analysis in ALDH1A1 KD cells of H929 and 8226 cells revealed that retinoic acid receptor, and retinoid X receptor agonists/antagonists pathways were enriched in both cell line; C. Knock down ALDH1A1caused downregulation of RAR proteins while re-introduced ALDH1A1 into ALDH1A1-KD cells increased their expressions; D . RA salvage restore the ADCC of Dara against MM cells; E . the RAR protein level was significantly increased determined by western blot after RA supplement; (F) mRNA levels of CD38 determined by RT-qPCR; (G) protein levels of CD38 determined by western blot. **, p < 0.01, ***, p < 0.001 C-G, student t test was adopted KD: ALDH1A1 knockdown; Ctrl: control; KD-OE: over expression of ALDH1A1 in KD cells; RA: retinoic acid; RAR: retinoic acid receptor; ADCC: antibody-dependent cellular cytotoxicity

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: Activation Assay, Knockdown, Western Blot, Quantitative RT-PCR, Control, Over Expression

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Reduced ALDH1A1 expression in multiple myeloma cells increases resistance to daratumumab via downregulation of retinoic acid

doi: 10.1007/s00018-025-05891-7

Figure Lengend Snippet: ALDH1A1 inhibitor reduces the sensitivity of Dara in vivo through inhibiting the RA/RAR pathway. A and B . compared with vehicle control, Dara cause significantly reduction of xenograft tumors weight and tumor growth; C . inhibitor of ALDH1A 673 A remarkably reduce the protein level of CD38, ALDH1A1 and RAR, and RA complement significantly restore the protein level of CD38 and RAR. *, p < 0.05, **, p < 0.01, ***, p < 0.001 A-B, analysis of variance (ANOVA) was used for multi-group comparisons followed by Tukey’s post hoc test ( N = 6); C, analysis of variance (ANOVA) was used for multi-group comparisons followed by Tukey’s post hoc test ( N = 3) RA: retinoic acid; RAR: retinoic acid receptor

Article Snippet: The positive CD38 cells were labeled by APC anti-human CD38 antibody (E-AB-F1058E, Elabscience) or APC anti-mouse CD38 antibody (E-AB-F1193E, Elabscience) and analyzed by FACSCantoTM II flow cytometer.

Techniques: In Vivo, Control

Journal: Bioengineering

Article Title: Human Mesenchymal Stem Cells Overexpressing Interleukin 2 Can Suppress Proliferation of Neuroblastoma Cells in Co-Culture and Activate Mononuclear Cells In Vitro

doi: 10.3390/bioengineering7020059

Figure Lengend Snippet: Changes in peripheral blood mononuclear cell (PBMC) activation marker expression after cultivation in hADSCs and hADSCs-IL2 CM. PBMCs were plated in 35 mm non-treated dishes (2 × 10 6 cells per dish) in 2 mL of CM harvested from hADSCs-IL2, native hADSCs or fresh DMEM/F12. Activation of PBMCs populations was determined using flow cytometry after 72 h. ( a ) Number of CD3 + /HLA-DR + late-activated T-cells, %. ( b ) Number of CD3 + /CD25 + early-activated T-cells, %. ( c ) Number of CD3 + /CD56 + NKT-cells, %. ( d ) Number of CD8 + /CD38 + activated T-killers, %. ( e ) Dot-plot with CD3 − /CD56 + NK cells cultured in DMEM/F12, hADSC CM, hADSCs-IL2 CM. Each box represents the mean ± SD ( n = 3) of two biological replicates. * p < 0.05, ** p < 0.01, ns: no significance.

Article Snippet: All samples were stained with allophycocyanin (APC)-conjugated anti-human CD3 (#300312, BioLegend, San Diego, CA, USA) or Alexa Fluor 488-conjugated anti-human CD3 (#2324020, Sony Biotechnology, San Jose, CA, USA), Alexa Fluor 488-conjugated HLA-DR (#IM0463U, Beckman Coulter, USA), Alexa Fluor 647-conjugated anti-human CD56 (NCAM) (#2191564, BioLegend, San Diego, CA, USA), R-phycoerythrin (PE)-Cy7-conjugated anti-human CD25 (#2113060, Sony Biotechnology, San Jose, CA, USA), PE-conjugated anti-human CD8a (#300908, BioLegend, San Diego, CA, USA) and Alexa Fluor 488-conjugated anti-human CD38 (#2117515, Sony Biotechnology, San Jose, CA, USA).

Techniques: Activation Assay, Marker, Expressing, Flow Cytometry, Cell Culture