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A. thaliana XVE_mTCP3 ER 16hr vs. XVE_mTCP3 DMSO 16hr

ABSTRACT: Cell expansion is crucial for organ morphogenesis in multicellular organisms. Plant cells are surrounded by rigid cell walls and turgor pressure drives cell expansion at a rate that is limited by the mechanical property of cell wall. The regulation of cell wall-loosening underlies the mechanism of cell expansion, but the integration of this regulation with organ morphogenesis is still elusive. In this study, we demonstrate that TEOSINTE BRANCHED1, CYCLOIDEA, PROLIFERATING CELL NUCLEAR ANTIGEN BINDING FACTORs (TCPs) transcription factors integrate cell wall loosening and organ morphogenesis in Arabidopsis thaliana. We observed that TCPs stimulated cell expansion and exaggerated elongation of hypocotyls. The atomic force microscopy analysis demonstrated that TCPs decreased the stiffness of cells, indicating the roles of TCPs to regulate the mechanical properties of cell wall. The gene expression analyses found that TCPs regulated the genes for both cell wall loosening and apoplast acidification such as SMALL AUXIN UP RNAs (SAURs). Because apoplast acidification induces cell wall loosening, we conducted western blot and pharmacological analyses and revealed that TCPs activated a plasma membrane-localized H+-ATPase responsible for apoplast acidification. Monitoring the apoplast pH at cellular resolutions indicated that TCPs stimulated apoplast acidification. Ectopic expression of a SAUR gene in a sextuple tcp mutant substantially recovered the hypocotyl morphology of the tcp mutant, providing the genetic evidence of the TCP-mediated SAUR regulation. Altogether, we demonstrate that TCPs govern cell wall loosening for cell expansion and pave the way to modify plant architectures through the regulation of cell wall loosening.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE271710 | GEO | 2025/09/23

REPOSITORIES: GEO

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Project description:Viral infections have a major impact on morbidity and mortality of immunosuppressed solid organ transplant (SOT) patients because of missing or failure of adequate pharmacologic antiviral treatment. Adoptive antiviral T-cell therapy (AVTT), regenerating disturbed endogenous T-cell immunity, emerged as an attractive alternative approach to combat severe viral complications in immunocompromised patients. AVTT is successful in patients after hematopoietic stem cell transplantation where T-cell products (TCPs) are manufactured from healthy donors. In contrast, in the SOT setting TCPs are derived from/applied back to immunosuppressed patients. We and others demonstrated feasibility of TCP generation from SOT patients and first clinical proof-of-concept trials revealing promising data. However, the initial efficacy is frequently lost long-term, because of limited survival of transferred short-lived T-cells indicating a need for next-generation TCPs. Our recent data suggest that Rapamycin treatment during TCP manufacture, conferring partial inhibition of mTOR, might improve its composition. The aim of this study was to confirm these promising observations in a setting closer to clinical challenges and to deeply characterize the next-generation TCPs. Using cytomegalovirus (CMV) as model, our next-generation Rapamycin-treated (Rapa-)TCP showed consistently increased proportions of CD4+ T-cells as well as CD4+ and CD8+ central-memory T-cells (TCM). In addition, Rapamycin sustained T-cell function despite withdrawal of Rapamycin and re-stimulation, showed superior T-cell viability and resistance to apoptosis, stable metabolism upon activation, preferential expansion of TCM, partial conversion of other memory T-cell subsets to TCM and increased clonal diversity. On transcriptome level, we observed a gene expression profile denoting long-lived early memory T-cells with potent effector functions. Furthermore, we successfully applied the novel protocol for the generation of Rapa-TCPs to 19/19 SOT patients in a comparative study, irrespective of their history of CMV reactivation. Moreover, comparison of paired TCPs generated before/after transplantation did not reveal inferiority of the latter despite exposition to maintenance immunosuppression post-SOT. Our data imply that the Rapa-TCPs, exhibiting longevity and sustained T-cell memory, are a reasonable treatment option for SOT patients. Based on our success to manufacture Rapa-TCPs from SOT patients under maintenance immunosuppression, now, we seek ultimate clinical proof of efficacy in a clinical study.

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A. thaliana XVE_mTCP3 ER 16hr vs. XVE_mTCP3 DMSO 16hr

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