ArticleInternational journal of molecular sciences2026
DTT-Induced Transcriptome Profiling Reveals Divergent ER Proteostasis and Stress-Adaptation Programs in CHO-S/HB8 and CHO-4BGD/DUL Producer Clones.
Article in International journal of molecular sciences, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Chinese hamster ovary (CHO) producer cells differ in their capacity to accommodate secretory and endoplasmic reticulum (ER) stress, but how producer and host-cell backgrounds are associated with ER-proteostasis responses remains poorly understood. We compared 24 h dithiothreitol (DTT)-induced reductive ER stress responses in two clonal producers: CHO-S-derived HB8 secreting human chorionic gonadotropin and apoptosis-resistant CHO-4BGD-derived DUL secreting a GLP-1-Fc fusion protein with similar specific productivities. Responses were analyzed by strand-specific RNA-seq, Xbp1-splicing RT-PCR, immunoblotting, functional enrichment, and curated-module analysis. Analysis of the complete three-replicate dataset identified 299 DTT-responsive genes in HB8 and 877 in DUL, demonstrating a broader transcriptional response in the DUL clone. Analysis after exclusion of the atypical HB8#3 matched pair yielded 404 and 1019 differentially expressed genes, respectively, and was used for detailed sensitivity analysis. Both producers showed ISR/ATF4/CHOP-associated transcriptional activation and Xbp1 mRNA splicing, whereas changes in total Xbp1 and Hspa5/BiP transcript abundance were limited. BiP and CHOP accumulation was detected at the protein level, while phospho-eIF2α and ATF6 responses were variable. Baseline RNA-seq data comparison revealed extensive transcriptional divergence between HB8 and DUL. HB8 showed higher expression of several classical ER folding/redox factors, whereas DUL showed higher expression of selected ISR-, quality-control- and stress-survival-associated genes. DUL additionally displayed broader vesicle/endocytic and amino-acid/glutathione-related remodeling. Thus, the producers occupy distinct ER-proteostasis states, and DUL mounts a broader, but not uniformly stronger, canonical UPR response to reductive ER stress.
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