Pleiotropic Functions Research Articles

Functional validation of a novel STAT3 'variant of unknown significance' identifies a new case of STAT3 GOF Syndrome and reveals broad immune cell defects.

STAT3 orchestrates crucial immune responses through its pleiotropic functions as a transcription factor. Patients with germline monoallelic dominant negative or hypermorphic STAT3 variants, who present with immunodeficiency and/or immune dysregulation, have revealed the importance of balanced STAT3 signaling in lymphocyte differentiation and function, and immune homeostasis. Here, we report a novel missense variant of unknown significance in the DNA binding domain of STAT3 in a patient who experienced hypogammaglobulinemia, lymphadenopathy, hepatosplenomegaly, immune thrombocytopenia, eczema and enteropathy over a 35-year period. In vitro demonstration of prolonged STAT3 activation due to delayed de-phosphorylation, and enhanced transcriptional activity, confirmed this to be a novel pathogenic STAT3 gain-of-function variant. Peripheral blood lymphocytes from this patient, and patients with confirmed STAT3 Gain-of-function Syndrome, were collected to investigate mechanisms of disease pathogenesis. B cell dysregulation was evidenced by a loss of class-switched memory B cells and a significantly expanded CD19hiCD21lo B cell population, likely influenced by a skewed CXCR3+ TFH population. Interestingly, unlike STAT3 dominant negative variants, cytokine secretion by activated peripheral blood STAT3 GOF CD4+ T cells and frequencies of Treg cells were intact, suggesting CD4+ T cell dysregulation likely occurs at sites of disease rather than the periphery. This study provides an in-depth case study in confirming a STAT3 gain-of-function variant and identifies lymphocyte dysregulation in peripheral blood of patients with STAT3 Gain-of-function Syndrome. Identifying cellular biomarkers of disease provide a flow cytometric based screen to guide validation of additional novel STAT3 gain-of-function variants as well as provide insights into putative mechanisms of disease pathogenesis.

The Balance Between the Natriuretic Peptides and the Renin-Angiotensin-Aldosterone System in the Preservation of Ideal Cardiovascular Health.

A healthy lifestyle plays a key role for maintaining the cardiovascular health (CVH) status and prevent cardiovascular disease occurrence. In fact, a healthy lifestyle was included in the AHA Cardiovascular Health score (Life's Simple 7 [LS7]), subsequently updated to Life's Simple 8 [LS8]. Apart from the importance of controlling conventional cardiovascular risk factors, increasing evidence supports the contributory role of cardiovascular hormones. Higher levels of natriuretic peptides (NPs) and lower levels of renin and aldosterone were significantly associated to CVH. NT-proBNP levels showed a direct relationship with CVH scores in large general Caucasian populations, being also a marker of CVH changes and a predictor of future adverse events. On the other hand, renin and aldosterone were inversely related to CVH scores. In contrast, the counter-regulatory angiotensins [Ang (1-7) acting through Mas receptor, Ang (1-9) acting through Angiotensin Type 2 receptor, and alamandine] strengthen the beneficial properties of NPs. This evidence can be explained by both the effects on systemic hemodynamic and possible pleiotropic local functions regulating different pathways involved in the maintenance of CVH. Based on the current evidence, circulating levels of NT-proBNP, renin and aldosterone may affect CVH in apparently asymptomatic individuals and represent additional markers of residual cardiovascular risk.

Histone H2A.Z Deacetylation and Dedifferentiation in Infarcted/Tip60-depleted Cardiomyocytes.

Myocardial infarction (MI) results in the loss of billions of cardiomyocytes (CMs), resulting in cardiac dysfunction. To re-muscularize injured myocardium, new CMs must be generated via renewed proliferation of surviving CMs. Approaches to induce proliferation of CMs after injury have been insufficient. Toward this end we are targeting the acetyltransferase Tip60, encoded by the Kat5 gene, based on the rationale that its pleiotropic functions combine to block CM proliferation at multiple checkpoints. We previously demonstrated that genetic depletion of Tip60 in a mouse model after MI reduces scarring, retains cardiac function, and activates the CM cell-cycle, although it remains unclear whether this culminates in the generation of daughter CMs. In order for pre-existing CMs in the adult heart to undergo proliferation, it has become accepted that they must first dedifferentiate, a process highlighted by loss of maturity, epithelial to mesenchymal transitioning (EMT), and reversion from fatty acid oxidation to glycolytic metabolism, accompanied by softening of the myocardial extracellular matrix (ECM). Based on recently published findings that Tip60 induces and maintains the differentiated state of hematopoietic stem cells and neurons via site-specific acetylation of the histone variant H2A.Z, we assessed levels of acetylated H2A.Z and dedifferentiation markers after depleting Tip60 in CMs post-MI. We report that genetic depletion of Tip60 from CMs after MI results in the near obliteration of acetylated H2A.Z in CM nuclei, accompanied by the altered expression of genes indicative of EMT induction, ECM softening, decreased fatty acid oxidation, and depressed expression of genes that regulate the TCA cycle. In accord with the possibility that site-specific acetylation of H2A.Z maintains adult CMs in a mature state of differentiation, CUT&Tag revealed enrichment of H2A.Zac K4/K7 in genetic motifs and in GO terms respectively associated with CM transcription factor binding and muscle development/differentiation. Along with our previous findings, these results support the notion that Tip60 has multiple targets in CMs that combine to maintain the differentiated state and prevent proliferation.

TGFβ signaling sensitizes MEKi-resistant human melanoma to targeted therapy-induced apoptosis

The TGFβ signaling pathway is known for its pleiotropic functions in a plethora of biological processes. In melanoma, TGFβ signaling promotes invasiveness and metastasis formation. However, its involvement in the response to therapy is controversial. While several studies have linked TGFβ signaling to elevated resistance to targeted therapy in melanoma, separate findings have indicated a favorable treatment response through TGFβ-mediated increase of cell death. We now found that the outcome of TGFβ signaling in the context of targeted therapy is dose dependent. Unlike low doses, high levels of TGFβ signal activation induce apoptosis upon simultaneous MAPK pathway inhibition, even in targeted therapy resistant melanoma cell lines. Using transcriptomic analyses, combined with genomic target identification of the critical TGFβ signaling effector SMAD4, we demonstrate that parallel activation of TGFβ signaling and MAPK pathway inhibition causes a complete switch of TGFβ target genes from promoting pro-invasive processes to fueling pro-apoptotic pathways. Investigations of underlying mechanisms identified a novel apoptosis-inducing gene signature. Functional validation of signature members highlighted a central role of the pro-apoptotic BCL2 family member BCL2L11 (BIM) in mediating apoptosis in this condition. Using a modified, synthetic version of the TGFB1 mRNA for intra-tumoral injections, we additionally showcase a potential therapeutic application of this treatment combination.

A brief history of insect neuropeptide and peptide hormone research.

This review briefly summarizes 50years of research on insect neuropeptide and peptide hormone (collectively abbreviated NPH) signaling, starting with the sequencing of proctolin in 1975. The first 25years, before the sequencing of the Drosophila genome, were characterized by efforts to identify novel NPHs by biochemical means, mapping of their distribution in neurons, neurosecretory cells, and endocrine cells of the intestine. Functional studies of NPHs were predominantly dealing with hormonal aspects of peptides and many employed ex vivo assays. With the annotation of the Drosophila genome, and more specifically of the NPHs and their receptors in Drosophila and other insects, a new era followed. This started with matching of NPH ligands to orphan receptors, and studies to localize NPHs with improved detection methods. Important advances were made with introduction of a rich repertoire of innovative molecular genetic approaches to localize and interfere with expression or function of NPHs and their receptors. These methods enabled cell- or circuit-specific interference with NPH signaling for in vivo assays to determine roles in behavior and physiology, imaging of neuronal activity, and analysis of connectivity in peptidergic circuits. Recent years have seen a dramatic increase in reports on the multiple functions of NPHs in development, physiology and behavior. Importantly, we can now appreciate the pleiotropic functions of NPHs, as well as the functional peptidergic "networks" where state dependent NPH signaling ensures behavioral plasticity and systemic homeostasis.

Beyond protein folding: The pleiotropic functions of PPIases in cellular processes and microbial virulence.

Peptidyl prolyl cis/trans isomerases (PPIases), a ubiquitously distributed superfamily of enzymes, associated with signal transduction, trafficking, assembly, biofilm formation, stress tolerance, cell cycle regulation, gene expression and tissue regeneration, is a key regulator of metabolic disorders and microbial virulence. This review assumes an integrative approach, to provide a holistic overview of the structural and functional diversity of PPIases, examining their conformational dynamics, cellular distribution, and physiological significance. We explore their intricate involvement in cellular processes and virulence modulation in both eukaryotic and prokaryotic systems. Additionally, we evaluate the potential of these molecular chaperones as drug targets and vaccine candidates, emphasizing their relevance in therapeutic development. By synthesizing recent findings and providing a broader perspective on these proteins, this review aims to enhance our understanding of their multifaceted roles in biology and their potential applications in medicine.

Identification of a novel prognostic signature for breast cancer derived from post-translational ubiquitin and ubiquitin-like modification-related genes.

Ubiquitin and ubiquitin-like (UUL) modifications play pleiotropic functions and are subject to fine regulatory mechanisms frequently altered in cancer. However, the comprehensive impact of UUL modification on breast cancer remains unclear. Transcriptomic and clinical data of breast cancer were downloaded from TCGA and GEO databases. Molecular subtyping of breast cancer was conducted using the NMF and CIBERSORT algorithms. Prognostic genes were identified via univariate, lasso and multivariate Cox regression analyses. Clinical pathological features, immune cell infiltration, immune therapeutic response and chemotherapy drug sensitivity were compared between groups using the Wilcoxon test. Survival analysis was performed using the Kaplan-Meier method and log-rank test. A total of 63 UUL modification-related genes were differentially expressed, with 29 up-regulated and 34 down-regulated genes. These genes were used to generate two UUL modification patterns that exhibited significant differences in prognostic features and immune cell infiltration. The UUL modification patterns were associated with 2038 differentially expressed genes that were significantly enriched in nuclear division, chromosome segregation, neuroactive ligand-receptor interaction, cell cycle, and other biological processes. Of these genes, 425 were associated with breast cancer prognosis, which enabled the classification of breast cancer into two clusters with significantly distinct prognoses. We developed a prognostic model, UULscore, which comprised nine genes and showed a significant correlation with partial immune cell infiltration. Furthermore, UULscore demonstrated potential predictive value in breast cancer overall survival prediction, immune therapeutic response, and chemotherapy drug sensitivity. UULscore, stage, radiotherapy, and chemotherapy were identified as independent prognostic factors for breast cancer. Based on these factors, a nomogram model was constructed, which demonstrated exceptional prognostic predictive performance. The present study identified two UUL modification-derived molecular subtypes in breast cancer, and have successfully constructed a risk-scoring model that holds potential value in prognosis, immune infiltration, immune therapeutic response, and chemotherapy sensitivity.

Allelic Variation in gtfB–gtfC Region of Natural Variant of Streptococcus mutans Without Biofilm Formation

Streptococcus mutans is primarily found in biofilms on tooth surfaces and is associated with the development of dental caries. S. mutans synthesizes water-insoluble glucan (WIG) using sucrose as a substrate, inducing the formation of three-dimensional biofilms. WIG is produced by glucosyltransferases (GTFs) encoded by the tandem and highly homologous gtfB and gtfC genes. Conversely, the homologous recombination of gtfB and gtfC readily happens, producing natural variants without WIG. These WIG− variants are thought to have ecologically pleiotropic functions; however, the molecular basis for their appearance is unclear. This study aimed to determine the sequence of the gtfB–gtfC regions of WIG− variants. We sequenced the gtfB–gtfC regions of 23 WIG− variants derived from S. mutans UA159 and revealed the presence of five alleles and four types of single homologous recombination patterns. Regardless of the allele type, the WIG− variants showed low biofilm formation and GTF activity. To the best of our knowledge, this is the first study to report the presence of alleles in WIG− variants. These findings provide important information for explaining the appearance of mechanisms in WIG− variants.

Characterization of Dnajc12 knockout mice, a model of hypodopaminergia.

Homozygous DNAJC12 c.79-2A>G (p. V27Wfs*14) loss-of-function mutations were first reported as a cause of young-onset Parkinson's disease. However, bi-allelic autosomal recessive pathogenic variants in DNAJC12 may lead to an alternative constellation of neurological features, including infantile dystonia, developmental delay, intellectual disability and neuropsychiatric disorders. DNAJC12 is understood to co-chaperone aromatic amino acid hydroxylases to foster the synthesis of biogenic amines. In vitro, we discover overexpressed DNAJC12 forms a complex with guanine triphosphate cyclohydrolase 1 (GCH1), the rate-limiting enzyme in the synthesis of tetrahydrobiopterin, a cofactor paramount for biogenic amines synthesis. We also confirm DNAJC12's interaction with tyrosine (TH) and tryptophan hydroxylase (TPH), which are rate-limiting enzymes for synthesis of biogenic amines dopamine (DA) and serotonin (5-HT). In-vitro knock-down of DNAJC12 with a siRNA destabilizes the DNAJC12-TH-GCH1 complex, reducing GCH1 levels, whereas reciprocal overexpression of both TH and GCH1 increases endogenous DNAJC12, alluding to the significance of modulating the DNAJC12-TH-GCH1 complex as a therapy for DNAJC12 and other biogenic amine disorders. We extend these investigations to a Cre-conditional knock-out mice (cDKO) in which loxP sites flanking Dnajc12 exon 2 enable its excision by cre-recombinase. With germline Cre expression, we have created a constitutive Dnajc12 knock-out (DKO). DKO mice exhibit reduced locomotion/ exploratory behavior at 3 months in automated open-field testing, accompanied by increased plasma phenylalanine which is a cardinal feature of patients with pathogenic DNAJC12 variants. In striatal tissue, total DA and 5-HT, their metabolites, and electrically-evoked DA release are all reduced. Biochemical alterations in synaptic proteins are also apparent, with enhanced phosphorylation of Th pSer31 and pSer40 reflecting biological compensation. Most immediately, cDKO and DKO mice present models to develop and refine therapeutic approaches for biogenic amines disorders, including dystonia and parkinsonism. They will also enable the pleiotropic functions of biogenic amines (including DA), usually synthesized in the brain or periphery, to be separated.

Divergent modulation of activated protein C pleiotropic functions by antibodies that differ by a single amino acid

AbstractActivated protein C (APC) is a pleiotropic plasma protease with diverse functions derived from its anticoagulant, anti-inflammatory, and cytoprotective activities. The selective uncoupling and/or modulation of these APC activities by antibodies may have therapeutic benefit in diseases such as traumatic bleeding, hemophilia, sepsis, and ischemia. TPP-26870 is an antibody that targets a nonactive site of APC for the selective modulation of APC activities. To optimize the potency of TPP-26870, variants with single amino acid mutation in the complementarity-determining regions (CDRs) were screened, and 21 variants with improved affinity constant were identified. Interestingly, the affinity maturation of TPP-26870 did not merely generate a panel of variants with higher potency in functional assays. Functional data demonstrated that the pleiotropic functions of APC were very sensitive to epitope-CDR interactions. Single amino acid mutations within the CDRs of TPP-26870 were sufficient to elicit divergent antagonistic and agonistic effects on the various APC functional activities. These include prolonged in vitro APC plasma half-life, increased inhibition of anticoagulant activity, and agonistic enhancement of histone H3 cleavage, while having less impact on protease-activated receptor 1 cleavage, compared with TPP-26870. This study illustrates that APC is highly sensitive to non–active site targeting that can lead to unpredictable changes in its activity profile of this pleiotropic enzyme. Furthermore, this study demonstrates the ability to modify APC functions to advance the potential development of APC-targeted antibodies as therapeutics for the treatment of diseases including trauma bleeding, hemophilia, ischemia, and sepsis.

Central role of squid gene during oocyte development in the Hemiptera Rhodnius prolixus

Oocyte polarity establishment is a conserved and crucial phenomenon for embryonic development. It relies on the precise spatial localization of maternal factors deposited during oocyte development, which is essential for establishing and maintaining cell polarity and subsequently specifying embryonic axes. The heterogeneous nuclear ribonucleoprotein (hnRNP) encoded by the squid (sqd) gene has been implicated in mRNA localization and embryonic axis establishment in Drosophila melanogaster. Comparative genomics allowed for the identification of a homologue in Rhodnius prolixus. In this study, we investigated the function of Rp-sqd during oogenesis and early embryonic development. We observed persistent expression of Rp-sqd during oocyte development, with localization in the cytoplasm of ovary germarium and growing oocytes in previtellogenic and vitellogenic stages. A Parental RNA interference (RNAi) experiment targeting Rp-sqd resulted in female sterility. The ovaries showed disrupted oocyte development, disarray of follicular epithelium, and affected nurse cells integrity. Immunostaining and microscopic techniques revealed microtubule disarray and a reduction in the presence of organelles in the trophic cords that connect the germarium with the oocytes. The Rp-sqd depletion impacted the transcript expression of maternal mRNAs involved in apoptosis, axis formation, oogenesis, and cytoskeleton maintenance, indicating a pleiotropic function of Rp-sqd during oogenesis. This study provides new insights into the genetic basis of R. prolixus oogenesis, highlighting the crucial role of Rp-sqd in oocyte development, fertility, and germarium integrity. These findings contribute to our understanding of insect developmental processes, provide a foundation for future investigations into reproduction, and reveal the regulatory mechanisms governing the process.

The role of nesfatin-1 in kidney diseases.

Nesfatin-1 is a recently discovered protein with a pleiotropic function on various organs, including kidneys. This molecule presents antiapoptotic, antihyperglycemic, antioxidative, and anorectic features. Available data regarding the role of nesfatin-1 in kidney function and diseases focuses on chronic kidney disease, acute kidney injury, blood pressure, and renal cell carcinoma. Various studies have shown that the levels of nesfatin-1 were increased in patients with diabetic kidney disease (DKD); therefore, it was suggested that nesfatin-1 might act as an early DKD marker. Furthermore, the potential protective function of nesfatin-1 against inflammation, oxidative stress, fibrosis, and apoptosis in kidney tissues was described in several studies. Alternatively, as reported in the literature, a positive correlation between blood pressure elevation and nesfatin-1 levels was noted. Moreover, nesfatin-1 might exert influence on renal cell carcinoma progression and invasion of cancerous cells. Nesfatin-1 shows considerable potential for acting as a prognostic marker or a defensive factor for kidney diseases; however, further investigation, especially in the pediatric population, is still required.

The role of STK11/LKB1 in cancer biology: implications for ovarian tumorigenesis and progression.

STK11 (serine-threonine kinase 11), also known as LKB1 (liver kinase B1) is a highly conserved master kinase that regulates cellular metabolism and polarity through a complex signaling network involving AMPK and 12 other AMPK-related kinases. Germline mutations in LKB1 have been causatively linked to Peutz-Jeghers Syndrome (PJS), an autosomal dominant hereditary disease with high cancer susceptibility. The identification of inactivating somatic mutations in LKB1 in different types of cancer further supports its tumor suppressive role. Deleterious mutations in LKB1 are frequently observed in patients with epithelial ovarian cancer. However, its inconsistent effects on tumorigenesis and cancer progression suggest that its functional impact is genetic context-dependent, requiring cooperation with other oncogenic lesions. In this review, we summarize the pleiotropic functions of LKB1 and how its altered activity in cancer cells is linked to oncogenic proliferation and growth, metastasis, metabolic reprogramming, genomic instability, and immune modulation. We also review the current mechanistic understandings of this master kinase as well as therapeutic implications with particular focus on the effects of LKB1 deficiency in ovarian cancer pathogenesis. Lastly, we discuss whether LKB1 deficiency can be exploited as an Achilles heel in ovarian cancer.

Phosphorylation by JNK switches BRD4 functions

Bromodomain 4 (BRD4), a key regulator with pleiotropic functions, plays crucial roles in cancers and cellular stress responses. It exhibits dual functionality: chromatin-bound BRD4 regulates remodeling through its histone acetyltransferase (HAT) activity, while promoter-associated BRD4 regulates transcription through its kinase activity. Notably, chromatin-bound BRD4 lacks kinase activity, and RNA polymerase II (RNA Pol II)-bound BRD4 exhibits no HAT activity. This study unveils one mechanism underlying BRD4's functional switch. In response to diverse stimuli, c-Jun N-terminal kinase (JNK)-mediated phosphorylation of human BRD4 at Thr1186 and Thr1212 triggers its transient release from chromatin, disrupting its HAT activity and potentiating its kinase activity. Released BRD4 directly interacts with and phosphorylates RNA Pol II, PTEFb, and c-Myc, thereby promoting transcription of target genes involved in immune and inflammatory responses. JNK-mediated BRD4 functional switching induces CD8 expression in thymocytes and epithelial-to-mesenchymal transition (EMT) in prostate cancer cells. These findings elucidate the mechanism by which BRD4 transitions from a chromatin regulator to a transcriptional activator.

Utilizing PRAME Expression and a Meta-Analytic Framework for iSALT to Explore Atypical Late-Onset Nevi of the Elderly and Their Relationship With Lentiginous and Nested Nevoid Melanomas.

In contrast to early-onset dysplastic nevi, late-onset atypical nevi of the elderly are more often precursors to distinctive nevoid melanomas. PReferentially expressed Antigen in MElanoma (PRAME) immunohistochemistry was applied to delineate the nevoid aspect of late-onset oncogenic nevoid pathway. Inducible Skin-Associated Lymphoid Tissue, regulatory T-cell mesenchymal hubs, has emerged as a translational tool and was used to define nevoid oncogenesis within a dynamic meta-analytic pathway. PRAME immunohistochemistry was applied after designating a histopathologic diagnosis. Late-onset atypical nested lentiginous nevus, lentiginous nested melanoma, and hypercellular nested nevoid melanoma were the diagnostic categories. A positive PRAME for melanoma was set at 75% percentage labeling.A wide-ranging published evidence-based database was incorporated to develop a meta-analytic framework for oncogenic nevogenesis. This combined inducible Skin-Associated Lymphoid Tissue incorporating the pleiotropic functions of regulatory T cells regulating immunity and gene regulatory epigenetics as principal modulators. Concordant-negative PRAME expression was present in 64 of 81 (79%) atypical nested lentiginous nevi, concordant-positive PRAME expression occurred in 54 of 75 (72%) nevoid lentiginous and nested melanomas, and 18 of 23 (78%) nevoid hypercellular nested melanomas. PRAME expression confirmed the existence of a late-onset oncogenic nevoid pathway that can be defined by histopathology. Subsequent meta-analysis data linked to the meta-analytic framework revealed that PRAME is an epigenetic surrogate antigen expressed because of repression of retinoic acid receptor signaling, preventing ligand-induced retinoic acid cellular differentiation, growth arrest, and apoptosis, and promoting melanoma growth and survival for melanomas. PRAME is only a single antigen within a highly complex dynamic framework that governs nevoid oncogenesis. Significantly, the retinoic acid/retinoic acid receptor complex has been shown to modulate the immunosuppressive arm of regulatory T cells underpinning immune tolerance and is pertinent to the broad framework but is not linked to PRAME expression in this arm.

LncRNA SNHG16 Inhibits Intracellular M. tuberculosis Growth Involving Cathelicidin Pathway, Autophagy, and Effector Cytokines Production.

Long noncoding small nucleolar RNA (LncRNA) host gene 16 (SNHG16) is associated with certain diseases, including cancers. However, its role and mechanism in Mycobacterium tuberculosis (Mtb) infection remain unclear. Here, we demonstrated that SNHG16 expression levels were suppressed in peripheral blood mononuclear cells (PBMCs) and CD14+ monocytes of tuberculosis (TB) patients. SNHG16 was up-regulated by acute Mtb infection of PBMCs from healthy control (HC) subjects. Such TB suppression of SNHG16 was consistent with an immunosuppressive-like state driven by IL-10 signaling as seen in TB patients. Notably, SNHG16 limited Mtb growth in macrophages/monocytes through autophagy and vitamin D receptor (VDR)-dependent cathelicidin (CAMP) antimicrobial pathways. Concurrently, SNHG16 was highly expressed in lymphocytes, including CD8+ and Vγ2 Vδ2 T-cell subsets in HCs. SNHG16 overexpression in lymphocytes allowed them to control Mtb infection in macrophages, and SNHG16 epigenetically increased the expression of anti-Mtb effector cytokines in lymphocytes by developing more accessible chromatin states in gene loci encoding IFN-γ, TNF-α, and Granzyme B. Furthermore, the adoptive transfer of SNHG16-overexpressing human PBMCs into Mtb-infected SCID mice conferred protective immunity against Mtb infection. Thus, SNHG16 drove the induction of pleiotropic effector functions that inhibited intracellular Mtb growth in vitro and in vivo, serving as an immunotherapy target in TB.

Phosphoproteome modulation by nucleoside diphosphate kinase affects photosynthesis & stress tolerance of Nostoc PCC 7120

Nucleoside diphosphate kinase (Ndk/NDK/NDPK) is known to possess pleiotropic functions, one of which is that as a protein kinase, and has been shown to be involved in stress tolerance in plants. To assess its role in the cyanobacterium Nostoc PCC 7120, which is hitherto unreported, recombinant strain overexpressing Ndk, Anndk+ was generated. Phosphoproteomic analysis of Anndk+ and its comparison with that of the vector control, AnpAM, revealed differential phosphorylation at S/T/Y sites of proteins belonging to varied functional groups, with over 17 % phosphoproteins involved in photosynthesis. A total of 177 phosphopeptides and 117 phosphoproteins were identified, including newly identified phosphopeptides in any cyanobacteria. Compared to AnpAM, the Anndk+ cells exhibited (i) lower photosynthetic efficiency and electron transport rate at low PAR (photosynthetically active radiation), (ii) no change in photochemical quenching across PAR, (iii) but distinct non-photochemical quenching [zero Y(NPQ) and high Y(NO) in Anndk+ and high Y(NPQ) and low (NO) in AnpAM], and (iv) increased tolerance to γ-radiation, oxidative, salt and DCMU stresses. The observed modulation of phosphoproteome linked to physiological changes upon overexpression of Ndk in Nostoc could be a combination of direct protein kinase activity of Ndk and/or indirectly through other protein kinases and phosphatases whose phosphorylation status is mediated by Ndk. This is the first report on a direct correlation between Ndk levels, phosphorylation status of proteins and stress tolerance in any cyanobacteria.

8553 GDF-15 Levels In Patients With Prostate Cancer On Androgen Deprivation Therapy

Abstract Disclosure: S.D. Jaramillo-Quiroz: Other; Self; VAPSHCS: Director Fund Research Fellowship/UW Abrass Family Fellowship Fund. CDMRP (PC170059).. L. Caeiro: None. L.J. Anderson: None. J.M. Garcia: None. Growth Differentiation Factor 15 (GDF-15) is a stress-induced cytokine known to induce anorexia and cachexia and a biomarker of biological aging, mitochondrial dysfunction, and worse hand grip strength and poor survival in different settings. Also, GDF-15 has pleiotropic functions at all stages of tumorigenicity and is suggested as a marker to differentiate BPH from PCa. Androgen Deprivation Therapy (ADT) is the standard treatment for advanced prostate cancer (PCa), but it also decreases muscle mass and function and Quality of life (QOL). The role and effects of GDF-15 in prostate cancer patients undergoing ADT are not well characterized and was explored in this longitudinal, 6-month, observational study that recruited men with (PCa) starting ADT (n=60, mean age 69). Body composition [dual energy X-ray absorptiometry (DEXA), body weight history], physical performance [handgrip strength (HGS), 6-minute walk test (6MWT), energy expenditure (indirect calorimetry and actigraphy)], were assessed before, at 3 and 6 months after starting ADT. QoL was assessed by the European Organization for the Research and Treatment of Cancer Quality of Life Questionnaire (EORTC-QLQ-C30). GDF-15 was measured by ELISA. Mean differences performed with standard error of the mean, repeated measures ANOVA and Tukey’s Post Hoc test. Correlations were assessed by Spearman’s. After 6 months of ADT, fat mass was increased (mean difference: 1.14 ±0.38, p=0.0029), and lean muscle mass (mean difference: -1.214 ± 0.38, p=0.005), appendicular lean mass (ALM, mean difference: -0.879 ± 0.19, p=<0.0001). HGS (mean difference: -2.927 ± 0.59, p=<0.0001), and VO2 max decreased (mean difference -3.31 ± 0.83, p=0.0007). Baseline GDF-15 (mean 1456.59 ± 1022.80) was correlated with the changes in weight (r: -0.38, p=0.01), BMI (r: -0.33, p=0.01), ALM (r: -0.29, p=0.04), and fatigue (r: -0.38, p=0.004). As shown, baseline GDF-15 was inversely correlated with changes in weight, appendicular lean mass, and fatigue in subjects undergoing ADT, highlighting the possibility of its role as a biomarker and therapeutic target of body composition and patient-reported outcomes in this setting. Presentation: 6/1/2024

In silico Assessment of Phytochemicals from Selected Plants as Prospective TGF‐β1 Inhibitors for Prostate Cancer Therapy

AbstractTransforming growth factor β1 (TGF‐β) is a cytokine with pleiotropic biological functions. Recently, its signaling pathway has been highlighted for its implicative paradoxical roles in prostate cancer (PCa). Suppressing downstream effects of this pathway by interfering with receptor complex formation through inhibition of the TGF‐β1 leads to its antitumor effects, illuminating the TGF‐β1 as a viable therapeutic target for PCa. Our compound library—established by a literature‐based approach that identified phytochemicals with published evidence against the TGF‐β1—was screened by employing molecular docking, density functional theory (DFT), and molecular dynamic (MD) simulations to identify TGF‐β1 inhibitors. Eight of the 24 phytochemicals docked from our compound library had a good binding affinity (ranging from −11.7 to −10 kcal/mol) to the TGF‐β1 (PDB: 1PY5). The phytochemicals displayed good stability and reactivity as revealed by the DFT analysis and a desirable pharmacokinetic profile. The top four phytochemical complexes with high binding energies maintained stability throughout the 100 ns simulation. Qualitative studies on the drug repurposing attributes of bisindolylmaleimide, flavopiridol, baicalin, and gefitinib as inhibitors of TGF‐β1 are recommended; most importantly, suggest further wet‐lab studies to corroborate these phytochemicals—SB 202190, SB 203580, silymarin, and cryptotanshinone—in TGF‐β1 targeted drug development.

The Functions and Application Prospects of Hepatocyte Growth Factor in Reproduction.

Hepatocyte growth factor (HGF) is expressed in multiple systems and mediates a variety of biological activities, such as mitosis, motility, and morphogenesis. A growing number of studies have revealed the expression patterns and functions of HGF in ovarian and testicular physiology from the prenatal to the adult stage. HGF regulates folliculogenesis and steroidogenesis by modulating the functions of theca cells and granulosa cells in the ovary. It also mediates somatic cell proliferation and steroidogenesis, thereby affecting spermatogenesis in males. In addition to its physiological effects on the reproductive system, HGF has shown advantages in preclinical studies over recent years for the treatment of male and female infertility, particularly in women with premature ovarian insufficiency. This review aims to summarize the pleiotropic functions of HGF in the reproductive system and to provide prospects for its clinical application.