Reversal Of Phenotype Research Articles

Taste receptor type 1 member 3 in osteoclasts regulates osteoclastogenesis via detection of glucose.

The taste system extends beyond the oral cavity, with various taste receptors found in extraoral organs. Mice deficient in the taste receptor type 1 (TAS1R) family member, TAS1R3, and fed a high-fat, high-sugar diet showed high bone mass without altering food consumption. However, the underlying mechanisms, including the cell types responsible for TAS1R3 expression, remain unclear. Here, we demonstrate the expression and function of TAS1R3 in osteoclasts, which are responsible for bone resorption. The expression of Tas1r3 but not Tas1r1 or Tas1r2, is evoked during osteoclast differentiation. Osteoclastogenesis-related genes were downregulated in TAS1R3-deficient mice, whereas the opposite phenotypes were elicited by TAS1R3 overexpression. Contrary to the common heterodimerization with TAS1R1 or TAS1R2, TAS1R3 formed a homodimer that functioned to detect glucose, enhance p38 phosphorylation, and induce osteoclastogenesis. These results provide novel insights into the role of TAS1R3 in bone metabolism and suggest that TAS1R3 may be a viable target for therapeutic agents in bone metabolic diseases.

KCNJ15 inhibits chemical-induced lung carcinogenesis and progression through GNB1 mediated Hippo pathway.

Polycyclic aromatic hydrocarbons (PAHs) have been regarded as important environmental carcinogens that can cause lung cancer. However, the underlying epigenetic mechanism during PAHs-induced lung carcinogenesis has remained largely unknown. Previously, we screened some novel epigenetic regulatory genes during 3-methylcholanthrene (3-MCA)-induced lung carcinogenesis, including the potassium inwardly rectifying channel subfamily J member 15 (KCNJ15) gene. This study aimed to investigate the expression regulation, function, and mechanism of KCNJ15 through database analysis, malignant transformed cell model, and xenograft tumor models. We found that KCNJ15 was remarkably under-expressed during lung carcinogenesis and progression. Elevated levels of DNA methylation were associated with a gradual decrease of KCNJ15 expression in 3-MCA-induced malignantly transformed HBE cells. High expression of KCNJ15 was strongly correlated with good survival prognosis in lung cancer patients. KCNJ15 overexpression significantly inhibited the growth, invasion, and migration of lung cancer cells both in vitro and in vivo. Conversely, knockdown of KCNJ15 resulted in an opposite phenotype. Moreover, KCNJ15 inhibited the Hippo pathway by activating YAP phosphorylation and inhibiting YAP expression. Mechanistically, there was a significant protein-protein interaction between KCNJ15 and the G protein subunit beta 1 (GNB1). GNB1 overexpression partially restored the inhibition of Hippo pathway regulated by KCNJ15. In summary, our data demonstrated that KCNJ15, as a novel epigenetic silencing tumor suppressor, regulates cell growth, invasion, and migration by binding to GNB1 protein mediating the Hippo-YAP signaling pathway during chemical-induced lung carcinogenesis and progression. It provides novel insights into epigenetic regulation mechanism during carcinogenesis induced by environmental pollutants.

Lipid Levels Increase to the Normal Range After Nonmyeloablative Hematopoietic Cell Transplantation for Sickle Cell Disease.

Individuals with sickle cell disease (SCD) have a unique type of dyslipidemia characterized by low total cholesterol (TC), low low-density lipoprotein cholesterol (LDL-c), low high-density lipoprotein cholesterol (HDL-c), and normal triglycerides (TG). This lipid state is theorized to be cardioprotective against atherosclerosis. In SCD, hematopoietic cell transplant (HCT) offers a potentially curative therapy. Long-term survivors of HCT for hematologic malignancies are at increased risk for dyslipidemia and atherosclerosis long-term. The effects of HCT on SCD dyslipidemia are unknown. This retrospective cohort study characterizes lipid profiles at baseline and after nonmyeloablative allogeneic HCT for SCD. We analyzed data from 116 patients after nonmyeloablative HLA-matched sibling or haploidentical HCT for SCD at the NIH from 2009 to 2021. TC, HDL-c, LDL-c, and TG were collected pre-HCT, 1-year post-HCT, and annually thereafter. Data were analyzed using linear generalized estimating equation regression modeling. Successful HCT was associated with a rise in TC, LDL-c, and HDL-c and a decline in TG post-HCT. After HCT, previously low lipid levels increased to the normal range. These changes occurred within the first year of HCT and were maintained thereafter. In patients with graft failure, TC and LDL-c levels remain unchanged from their pre-HCT baseline. Sirolimus use for graft versus host disease prophylaxis was associated with higher TG levels. These findings suggest that SCD dyslipidemia resolves with reversal of the SCD phenotype. The normalization of lipid parameters suggests SCD patients are not at increased risk for atherosclerosis after successful HCT compared to their peers; further studies with longer follow-up are required.

Genome-wide characterization of RsHDAC gene members unravels a positive role of RsHDA9 in thermotolerance in radish (Raphanus sativus L.).

Radish is an economically important root vegetable crop worldwide. Histone deacetylases (HDACs), one of the most important epigenetic regulators, play prominent roles in plant growth and development as well as abiotic stress responses. Nevertheless, the systematical characterization and critical roles of HDAC gene members in thermogenesis remains elusive in radish. Herein, a total of 21 RsHDAC genes were identified from the radish genome. Among them, two RsSRTs, six RsHDTs and 13 RsHDAs were classified into the SIR2, HD2 and RPD3/HDA1subfamily, respectively. The RNA-seq analysis indicated that three RsHDAs (RsHDA6.1, RsHDA6.2 and RsHDA19) and five RsHDTs exhibited high expression in vascular cambium of radish taproot. Both the RsHDT3 and RsHDA9 showed dramatically up-regulated expression under heat, salt and three heavy metals treatments. Moreover, the transient LUC reporter assay revealed that the promoter activity of the nucleus-localized RsHDA9 was intensely induced by heat stress. Intriguingly, overexpression of RsHDA9 promoted thermotolerance via enhancing proline accumulation and scavenging of reactive oxygen species in radish cotyledons, whereas the supplement of trichostatin A (TSA) led to the opposite phenotype. Notably, RsWRKY26 bound to the RsHDA9 promoter and activated its transcription to achieve enhancing thermotolerance in radish. Collectively, these findings would facilitate deciphering molecular mechanism underlying RsHDA9-mediated regulatory network of thermogenesis in radish.

The SlWRKY42-SlMYC2 module synergistically enhances tomato saline-alkali tolerance by activating the jasmonic acid signaling and spermidine biosynthesis pathway.

Tomato (Solanum lycopersicum) is an important crop but frequently experiences saline-alkali stress. Our previous studies have shown that exogenous spermidine (Spd) could significantly enhance the saline-alkali resistance of tomato seedlings, in which a high concentration of Spd and jasmonic acid (JA) exerted important roles. However, the mechanism of Spd and JA accumulation remains unclear. Herein, SlWRKY42, a Group II WRKY transcription factor, was identified in response to saline-alkali stress. Overexpression of SlWRKY42 improved tomato saline-alkali tolerance. Meanwhile, SlWRKY42 knockout mutants, exhibited an opposite phenotype. RNA-sequencing data also indicated that SlWRKY42 regulated the expression of genes involved in JA signaling and Spd synthesis under saline-alkali stress. SlWRKY42 is directly bound to the promoters of SlSPDS2 and SlNHX4 to promote Spd accumulation and ionic balance, respectively. SlWRKY42 interacted with SlMYC2. Importantly, SlMYC2 is also bound to the promoter of SlSPDS2 to promote Spd accumulation and positively regulated saline-alkali tolerance. Furthermore, the interaction of SlMYC2 with SlWRKY42 boosted SlWRKY42's transcriptional activity on SlSPDS2, ultimately enhancing the tomato's saline-alkali tolerance. Overall, our findings indicated that SlWRKY42 and SlMYC2 promoted saline-alkali tolerance by the Spd biosynthesis pathway. Thus, this provides new insight into the mechanisms of plant saline-alkali tolerance responses triggered by polyamines (PAs).

Rapid and scalable personalized ASO screening in patient-derived organoids

Personalized antisense oligonucleotides (ASOs) have achieved positive results in the treatment of rare genetic disease1. As clinical sequencing technologies continue to advance, the ability to identify patients with rare disease harbouring pathogenic genetic variants amenable to this therapeutic strategy will probably improve. Here we describe a scalable platform for generating patient-derived cellular models and demonstrate that these personalized models can be used for preclinical evaluation of patient-specific ASOs. We describe protocols for delivery of ASOs to patient-derived organoid models and confirm reversal of disease-associated phenotypes in cardiac organoids derived from a patient with Duchenne muscular dystrophy (DMD) with a structural deletion in the gene encoding dystrophin (DMD) that is amenable to treatment with existing ASO therapeutics. Furthermore, we designed novel patient-specific ASOs for two additional patients with DMD (siblings) with a deep intronic variant in the DMD gene that gives rise to a novel splice acceptor site, incorporation of a cryptic exon and premature transcript termination. We showed that treatment of patient-derived cardiac organoids with patient-specific ASOs results in restoration of DMD expression and reversal of disease-associated phenotypes. The approach outlined here provides the foundation for an expedited path towards the design and preclinical evaluation of personalized ASO therapeutics for a broad range of rare diseases.

P0251 Inter-rater disagreements in applying the Montreal classification for Crohn’s disease: The Five-Nations Survey Study

Abstract Background The Montreal classification has been widely used in Crohn’s disease (CD) since 2005 to categorize patients by the age of onset (A), disease location (L), behavior (B), and upper gastrointestinal tract and perianal involvement. With evolving management paradigms in CD, we aimed to assess the performance of gastroenterologists in applying the Montreal classification. Methods An online survey was conducted among participants in an international educational conference on inflammatory bowel diseases (IBD). Participants classified 20 theoretical CD cases using the Montreal classification. Agreement rates with the IBD board (three expert gastroenterologists whose consensus rating was considered the gold standard) were calculated for gastroenterologist specialists and fellows/specialists with ≤2 years of clinical experience. A majority vote<75% among participants was considered a notable disagreement. The same cases were classified using three large language models (LLMs), ChatGPT-4, Claude-3, and Gemini-1.5, and assessed for agreement with the board and gastroenterologists. Fleiss Kappa was used to assess within-group agreement. Results Thirty-eight participants from five countries completed the survey. In defining the Montreal classification as a whole, specialists (21/38 [55%]) had a higher agreement rate with the board compared to fellows/young specialists (17/38 [45%]) [58% vs. 49%, p=0.012] and to LLMs (58% vs. 18%, p<0.001). Disease behavior classification was the most challenging, with 76% agreement among specialists and fellows/young specialists and 48% among LLMs compared to the IBD board. Regarding disease behavior within-group agreement was moderate (specialists: k=0.522, fellows/young specialists: k=0.532, LLMs: k=0.577; p<0.001 for all). Notable points of disagreement included: defining disease behavior concerning obstructive symptoms, assessing disease extent via video capsule endoscopy, and evaluating treatment-related reversibility of the disease phenotype. Conclusion There is significant inter-rater disagreement in applying the Montreal classification, particularly for disease behavior in CD. Improved education or revisions to phenotype criteria may be needed to enhance consensus on the Montreal classification.

HVEM as a Tumor-Intrinsic Regulator in Non-Small Cell Lung Cancer: Suppression of Metastasis via Glycolysis Inhibition and Modulation of Macrophage Polarization.

Herpes virus entry mediator (HVEM) is a novel costimulatory molecule which mediates stimulatory or inhibitory signals in immune responses which makes it an attractive target in cancer therapeutics. However, the role of tumor cell intrinsic HVEM on tumor biology remains largely unknown. In this study, We demonstrated that CK+HVEM+ tumor correlates with better survival using Multiplex immuno histochemistry (mIHC) in Human Lung Adenocarcinoma Tissue microarray. Next, we showed that HVEM knockdown promoted NSCLC cell invasion and metastasis in vitro whereas exhibited no effect on proliferation. Conversely, HVEM overexpression results in the opposite phenotype. Meanwhile, the conclusion were further confirmed in vivo experiment that overexpression of HVEM reduced the invasion and metastasis of NSCLC whereas no effect on tumor mass. Besides, vivo experiment showed that M1 TAMs in the HVEM overxrpression group was increased and the proportion of M2 macrophages was decreased compared to the vector group. Mechanistically, The C-terminal 228-283 amino acid segment of HVEM protein interacts with the N-terminal 1-383 amino acid segment of MPRIP protein, inhibiting its downstream glycolysis signaling pathway and suppressing NSCLC cells progression. In addition, macrophage coculture assay suggested that HVEM overexpression inhibited M2 macrophage polarization through GM-CSF/GM-CSFRα axis. In summary, our study has demonstrated that tumor cell intrinsic HVEM is a potential tumour metastasis suppressor, which may serve as a potential target for immunotherapy.

Inter-Rater Disagreements in Applying the Montreal Classification for Crohn's Disease: The Five-Nations Survey Study.

The Montreal classification has been widely used in Crohn's disease since 2005 to categorize patients by the age of onset (A), disease location (L), behavior (B), and upper gastrointestinal tract and perianal involvement. With evolving management paradigms in Crohn's disease, we aimed to assess the performance of gastroenterologists in applying the Montreal classification. An online survey was conducted among participants at an international educational conference on inflammatory bowel diseases. Participants classified 20 theoretical Crohn's disease cases using the Montreal classification. Agreement rates with the inflammatory bowel diseases board (three expert gastroenterologists whose consensus rating was considered the gold standard) were calculated for gastroenterologist specialists and fellows/specialists with ≤2years of clinical experience. A majority vote <75% among participants was considered a notable disagreement. The same cases were classified using three large language models (LLMs), ChatGPT-4, Claude-3, and Gemini-1.5, and assessed for agreement with the board and gastroenterologists. Fleiss Kappa was used to assess within-group agreement. Thirty-eight participants from five countries completed the survey. In defining the Montreal classification as a whole, specialists (21/38 [55%]) had a higher agreement rate with the board compared to fellows/young specialists (17/38 [45%]) (58% vs. 49%, p=0.012) and to LLMs (58% vs. 18%, p<0.001). Disease behavior classification was the most challenging, with 76% agreement among specialists and fellows/young specialists and 48% among LLMs compared to the inflammatory bowel diseases board. Regarding disease behavior, within-group agreement was moderate (specialists: k=0.522, fellows/young specialists: k=0.532, LLMs: k=0.577; p<0.001 for all). Notable points of disagreement included: defining disease behavior concerning obstructive symptoms, assessing disease extent via video capsule endoscopy, and evaluating treatment-related reversibility of the disease phenotype. There is significant inter-rater disagreement in applying the Montreal classification, particularly for disease behavior in Crohn's disease. Improved education or revisions to phenotype criteria may be needed to enhance consensus on the Montreal classification.

HDC1 Promotes Primary Root Elongation by Regulating Auxin and K+ Homeostasis in Response to Low-K+ Stress.

Plants frequently encounter relatively low and fluctuating potassium (K+) concentrations in soil, with roots serving as primary responders to this stress. Histone modifications, such as de-/acetylation, can function as epigenetic markers of stress-inducible genes. However, the signaling network between histone modifications and low-K+ (LK) response pathways remains unclear. This study investigated the regulatory role of Histone Deacetylase Complex 1 (HDC1) in primary root growth of Arabidopsis thaliana under K+ deficiency stress. Using a hdc1-2 mutant line, we observed that HDC1 positively regulated root growth under LK conditions. Compared to wild-type (WT) plants, the hdc1-2 mutant exhibited significantly inhibited primary root growth under LK conditions, whereas HDC1-overexpression lines displayed opposite phenotypes. No significant differences were observed under HK conditions. Further analysis revealed that the inhibition of hdc1-2 on root growth was due to reduced apical meristem cell proliferation rather than cell elongation. Notably, the root growth of hdc1-2 showed reduced sensitivity compared to WT after auxin treatment under LK conditions. HDC1 may regulate root growth by affecting auxin polar transport and subsequent auxin signaling, as evidenced by the altered expression of auxin transport genes. Moreover, the organ-specific RT-qPCR analyses unraveled that HDC1 negatively regulates the expression of CBL-CIPK-K+ channel-related genes such as CBL1, CBL2, CBL3, AKT1, and TPK1, thereby establishing a molecular link between histone deacetylation, auxin signaling, and CBLs-CIPKs pathway in response to K+ deficiency.

Revealing hidden knowledge in amnestic mice.

Alzheimer's disease (AD) is a form of dementia in which memory and cognitive decline is thought to arise from underlying neurodegeneration. These cognitive impairments, however, are transient when they first appear and can fluctuate across disease progression. Here, we investigate the neural mechanisms underlying fluctuations of performance in amnestic mice. We trained APP/PS1+ mice on an auditory go/no-go task that dissociated learning of task contingencies (knowledge) from its more variable expression under reinforcement (performance). APP/PS1+ exhibited significant performance deficits compared to control mice. Using large-scale two-photon imaging of 6,216 excitatory neurons in 8 mice, we found that auditory cortical networks were more suppressed, less selective to the sensory cues, and exhibited aberrant higher-order encoding of reward prediction compared to control mice. A small sub-population of neurons, however, displayed the opposite phenotype, reflecting a potential compensatory mechanism. Volumetric analysis demonstrated that deficits were concentrated near Aβ plaques. Strikingly, we found that these cortical deficits were reversed almost instantaneously on probe (non-reinforced) trials when APP/PS1+ performed as well as control mice, providing neural evidence for intact stimulus-action knowledge despite variable ongoing performance. A biologically-plausible reinforcement learning model recapitulated these results and showed that synaptic weights from sensory-to-decision neurons were preserved (i.e. intact stimulus-action knowledge) despite poor performance that was due to inadequate contextual scaling (i.e. impaired performance). Our results suggest that the amnestic phenotype is transient, contextual, and endogenously reversible, with the underlying neural circuits retaining the underlying stimulus-action associations. Thus, memory deficits commonly observed in amnestic mouse models, and potentially at early stages of dementia in humans, relate more to contextual drivers of performance rather than degeneration of the underlying memory traces.

Prognostic Significance of S100A4 in Ovarian Clear Cell Carcinoma: Its Relation to Tumor Progression and Chemoresistance.

S100A4, a small calcium-binding protein, promotes metastasis in a variety of human malignancies, but little is known about its involvement in ovarian clear cell carcinoma (OCCC). Herein, we characterized the functional role of S100A4 in this tumor type. We analyzed immunohistochemical sections from 120 OCCC patients. OCCC cell lines in which S100A4 was knocked out (KO) or overexpressed were also used to study the protein's effects. Stable overexpression of S100A4 decreased the proliferation of OCCC cell lines (concomitant with more cells in G1 and fewer in the G2/M phase of the cell cycle). S100A4 overexpression also reduced susceptibility to cisplatin-induced apoptosis (probably due to an increased BCL2: BAX ratio), accelerated epithelial-mesenchymal transition (EMT)-related cell mobility, and enhanced cancer stem cell (CSC) properties (including increases in both spheroid formation and in the aldehyde dehydrogenase 1 (ALDH1)high population). In contrast, S100A4 KO generally induced the opposite phenotypes, although it did not affect migration capability. In clinical OCCC samples, high S100A4 expression was associated with a low frequency of cleaved poly-(ADP-ribose) polymerase 1-positive apoptotic cells, a reduced proliferative rate, and expression of high ALDH1 and vimentin levels. In addition, a high S100A4 score was a significant (but not independent) prognostic factor in OCCC. Our findings suggest that S100A4 may be an unfavorable prognostic factor in OCCC, as it accelerates tumor progression and promotes chemoresistance through the modulation of proliferation, susceptibility to apoptosis, and EMT/CSC properties.

Senolytic treatment attenuates immune cell infiltration without improving IAV outcomes in aged mice

AbstractAging is a major risk factor for poor outcomes following respiratory infections. In animal models, the most severe outcomes of respiratory infections in older hosts have been associated with an increased burden of senescent cells that accumulate over time with age and create a hyperinflammatory response. Although studies using coronavirus animal models have demonstrated that removal of senescent cells with senolytics, a class of drugs that selectively kills senescent cells, resulted in reduced lung damage and increased survival, little is known about the role that senescent cells play in the outcome of influenza A viral (IAV) infections in aged mice. Here, we tested if the aged mice survival or weight loss IAV infections could be improved using three different senolytic regimens. We found that neither dasatinib plus quercetin, fisetin, nor ABT‐263 improved outcomes. Furthermore, both dasatanib plus quercetin and fisetin treatments further suppressed immune infiltration than aging alone. Additionally, our data show that the short‐term senolytic agents do not reduce senescent markers in our aged mouse model. These findings suggest that acute senolytic treatments do not universally reverse aging related immune phenotype against all respiratory viral infections.

Cotton RLP6 Interacts With NDR1/HIN6 to Enhance Verticillium Wilt Resistance via Altering ROS and SA.

Cotton Verticillium wilt (VW) is often a destructive disease that results in significant fibre yield and quality losses in Gossypium hirsutum. Transferring the resistance trait of Gossypium barbadense to G. hirsutum is optional but challenging in traditional breeding due to limited molecular dissections of resistance genes. Here, we discovered a species-diversified structural variation (SV) in the promoter of receptor-like protein 6 (RLP6) that caused distinctly higher expression level of RLP6 in G. barbadense with the SV than G. hirsutum without the SV. Functional experiments showed that RLP6 is an important regulator in mediating VW resistance. Overexpressing RLP6 significantly enhanced resistance and root growth, whereas the opposite phenotype appeared in RLP6-silenced cotton. A series of experiments indicated that RLP6 regulated reactive oxygen species (ROS) and salicylic acid (SA) signalling, which induced diversified defence-related gene expression with pathogenesis-related (PR) proteins and cell wall proteins enrichments for resistance improvement. These findings could be valuable for the transfer of the G. barbadense SV locus to improve G. hirsutum VW resistance in future crop disease resistance breeding.

Roles for the long non-coding RNA Pax6os1/PAX6-AS1 in pancreatic beta cell function.

Long non-coding RNAs (lncRNAs) are emerging as crucial regulators of beta cell function. Here, we show that an lncRNA-transcribed antisense to Pax6, annotated as Pax6os1/PAX6-AS1, was upregulated by high glucose concentrations in human as well as murine beta cell lines and islets. Elevated expression was also observed in islets from mice on a high-fat diet and patients with type 2 diabetes. Silencing Pax6os1/PAX6-AS1 in MIN6 or EndoC-βH1 cells increased several beta cell signature genes' expression. Pax6os1/PAX6-AS1 was shown to bind to EIF3D, indicating a role in translation of specific mRNAs, as well as histones H3 and H4, suggesting a role in histone modifications. Important interspecies differences were found, with a stronger phenotype in humans. Only female Pax6os1 null mice fed a high-fat diet showed slightly enhanced glucose clearance. In contrast, silencing PAX6-AS1 in human islets enhanced glucose-stimulated insulin secretion and increased calcium dynamics, whereas overexpression of the lncRNA resulted in the opposite phenotype.

Dissecting Causal Relationships Between Antihypertensive Drug, Gut Microbiota, and Type 2 Diabetes Mellitus and Its Complications: A Mendelian Randomization Study.

Limited research has investigated the impact of antihypertensive medications on type 2 diabetes mellitus (T2DM) and whether gut microbiome (GM) mediates this association. Thus, we conducted a two-sample Mendelian randomization (MR) analysis to estimate the potential impact of various antihypertensive drug target genes on T2DM and its complications. Genetic instruments for the expression of antihypertensive drug target genes were identified with expression quantitative trait loci (eQTL) in blood, which should be associated with systolic blood pressure (SBP). Sensitivity analysis, including reverse causality detection, horizontal pleiotropy, phenotype scanning, and Bayesian colocalization, was used to validate our findings. We performed a two-step MR to detect the mediating role of GM. A 1-standard deviation (SD) decrease of KCNJ11 (acting on arteriolar smooth muscle, e.g., Pinacidil) gene expression was associated with lower SBP of 1.12 (95% confidence interval [CI], 0.93-1.31) mmHg, and a decreased risk of diabetic retinopathy (odds ratio [OR], 0.63; 95% CI, 0.52-0.76). Similarly, a 1-SD decrease of SLC12A2 (genetically a proxy for diuretics, for example, Torasemide) gene expression was correlated with a reduced risk of T2DM (OR, 0.88; 95% CI, 0.83-0.92). Interestingly, this causal effect was influenced by a decrease in the gut microbiota abundance of the genus Ruminococcus (effect proportion = 11.2%). Colocalization supports these results (KCNJ11: 98% for diabetic retinopathy; SLC12A2: 99% for T2DM). Findings provide novel targets for the treatment of T2DM and its complications, emphasize the importance of KCNJ11 and SLC12A2 in future drug development, and highlight the significant mediating role of the genus Ruminococcus.

Role of biotin carboxyl carrier protein subunit 2 (BCCP2) in resistance to multiple stresses in Arabidopsis thaliana.

Abiotic stresses, including drought, salinity, and temperature extremes, are serious constraints to plant growth and agricultural development. These stresses that plants face in nature are often multiple and complex. Biotin carboxyl carrier protein subunit 2 (BCCP2) is one of the two subunits of biotin carboxyl carrier protein, which is a functional subunit of acetyl coenzyme A carboxylase, primarily studied for its role in fatty acid synthesis. In this study, we identified the expression pattern of AtBCCP2 under various stress conditions, including 4mM NaHCO₃, 2mM Na₂CO₃, 150mM NaCl, 300mM D-mannitol, 100μM ABA, 5mMH₂O₂, 4°C, and 37°C. It was determined that AtBCCP2 is positively regulated by NaHCO₃, Na₂CO₃, NaCl, and ABA, but negatively regulated by D-mannitol. Phenotypic experiment confirmed that the AtBCCP2 transgenic overexpression plants exhibited increased resistance to NaHCO₃, Na₂CO₃, NaCl, and ABA stresses, but more sensitive to drought stress simulated by D-mannitol. In contrast, mutant plants showed the opposite phenotypes. Additionally, AtBCCP2 transgenic overexpression plants demonstrated stronger antioxidant activity and lower MDA content under stresses such as NaHCO₃, Na₂CO₃, NaCl, and ABA, in contrast to the mutant plants. In response to D-mannitol simulated drought stress, AtBCCP2 transgenic overexpression plants showed lower antioxidant activity and higher MDA content, while mutant plants exhibited the opposite trend. In conclusion, this study provides a theoretical basis for understanding the role of AtBCCP2 in response to multiple stresses and contributes a new gene to the pool of those involved in abiotic stress responses.

Smad2 Cooperating with TGIF2 Contributes to EMT and Cancer Stem Cells Properties in Pancreatic Cancer via Co-Targeting SOX2.

The underlying mechanisms between cancer stem cells (CSC) and epithelial-mesenchymal transition (EMT) in pancreatic cancer (PC) remain unclear. In this study, we identified TGIF2 as a target gene of CSC using sncRNA and machine learning. TGIF2 is closely related to the expression of SOX2, EGFR, and E-cadherin, indicating poor prognosis. Mechanistically, TGIF2 promoted the EMT phenotype and CSC properties following the activation of SOX2, Slug, CD44, and ERGF/MAPK signaling, which were rescued by SOX2 silencing. TGIF2 silencing contributes to the opposite phenotype via SOX2. Notably, Smad2 cooperates with TGIF2 to co-regulate the SOX2 promoter, which in turn promotes EMT and CSC signaling by transactivating Slug and EGFR, respectively. The transactivation of EGFR/MAPK signaling by SOX2 promotes TGIF2 nuclear translocation, forming a positive feedback loop in vitro. Moreover, the interaction of TGIF2 and SOX2 with EGFR inhibitors promoted subcutaneous tumors and liver metastasis in vivo. Thus, the TGIF2/SOX2 axis contributes to CSC, EMT, and chemoresistance, providing a promising target for PC therapy.

Independent genetic strategies define the scope and limits of CDKL5 deficiency disorder reversal.

Cyclin-dependent kinase-like 5 (CDKL5) deficiency disorder (CDD) is a neurodevelopmental syndrome caused by mutations in the X-linked CDKL5 gene. The early onset of CDD suggests that CDKL5 is essential during development, but post-developmental re-expression rescues multiple CDD-related phenotypes in hemizygous male mice. Since most patients are heterozygous females, studies in clinically relevant female models are essential. Here, we systematically compare phenotype reversal across age and sex using two independent mouse models of CDD. We find that early re-activation of endogenous Cdkl5 in heterozygous females reverses most phenotypes, except working memory. Later re-expression improves several traits but has limited effects on cognitive function. Seizure prevention is more effective with early intervention in heterozygous females but becomes limited after seizure onset. These findings demonstrate the robust potential of CDKL5 re-expression to reverse CDD-related phenotypes in both sexes while underscoring the critical impact of age and disease stage in designing clinical trials.

COBRA-LIKE 9 modulates cotton cell wall development via regulating cellulose deposition.

Plant cell walls are complex and dynamic cellular structures critical for plant growth, development, physiology, and adaptation. Cellulose is one of the most important components of the cell wall. However, how cellulose microfibrils deposit and assemble into crystalline cellulose remains elusive. The COBRA-LIKE plant-specific protein family plays a vital role in modulating the deposition and orientation of cellulose microfibril in plant cell walls. Here, we investigate the role of GhCOBL9 in cotton (Gossypium hirsutum) fiber development, an ideal model for studying cell elongation and cell wall thickening. The expression period of GhCOBL9 is consistent with the thickening stage of the secondary wall of cotton fibers. Overexpression of GhCOBL9 results in increased cellulose content in the cell wall and produces shorter, thicker, and stronger fibers, while RNA interference (RNAi)-mediated downregulation of GhCOBL9 leads to the opposite phenotypes, indicating its crucial role in cell wall development. Subcellular localization and binding activity assays reveal that GhCOBL9 targets the cell wall and binds to crystalline cellulose with high affinity. Transcriptomic analysis of GhCOBL9 transgenic lines uncovers expression alterations in genes related to cellulose and monosaccharide biosynthesis. Furthermore, we identify a fasciclin-like arabinogalactan protein 9 (GhFLA9) as an interacting partner of GhCOBL9 to modulate cell wall development. Additionally, the R2R3-MYB transcription factor GhMYB46-5 activates GhCOBL9 expression by binding to the MYB46-responsive cis-regulatory element in the GhCOBL9 promoter. These findings broaden our knowledge of COBL function in modulating plant cell wall development.