Category Archives: BioFactors

Modulatory effects of point‐mutated IL‐32θ (A94V) on tumor progression in triple‐negative breast cancer cells

Posted on 2 September 2023 by

IL-32θ (A94V) inhibits phosphorylation of FAK and IκBα. IL-32θ (A94V) inhibits the expression and translocation of β-catenin by inhibiting phosphorylated FAK. Additionally, NF-κB is inhibited by IL-32θ (A94V) via the suppression of phosphorylated IκBα. Thus, IL-32θ (A94V) reduces migration, proliferation, and inflammation in breast cancer via the FAK-PI3K-GSK3 and NF-κB pathways.

Abstract

Breast cancer is a frequently diagnosed cancer and the leading cause of death among women worldwide. Tumor-associated macrophages stimulate cytokines and chemokines, which induce angiogenesis, metastasis, proliferation, and tumor-infiltrating immune cells. Although interleukin-32 (IL-32) has been implicated in the development and modulation of several cancers, its function in breast cancer remains elusive. Mutation of interleukin-32θ (IL-32θ) in the tissues of patients with breast cancer was detected by Sanger sequencing. RT-qPCR was used to detect the mRNA levels of inflammatory cytokines, chemokines, and mediators. The secreted proteins were detected using respective enzyme-linked immunosorbent assays. Evaluation of the inhibitory effect of mutant IL-32θ on proliferation, migration, epithelial–mesenchymal transition (EMT), and cell cycle arrest in breast cancer cells was conducted using MTS assays, migration assays, and Western blotting. A point mutation (281C>T, Ala94Val) was detected in IL-32θ in both breast tumors and adjacent normal tissues, which suppressed the expression of pro-inflammatory factors, EMT factors, and cell cycle related factors. Mutated IL-32θ inhibited the expression of inflammatory factors by regulating the NF-κB pathway. Furthermore, mutated IL-32θ suppressed EMT markers and cell cycle related factors through the FAK/PI3K/AKT pathway. It was inferred that mutated IL-32θ modulates breast cancer progression. Mutated IL-32θ (A94V) inhibited inflammation, EMT, and proliferation in breast cancer by regulating the NF-κB (p65/p50) and FAK-PI3K-GSK3 pathways.

Biochemical and cellular studies of three human 3‐phosphoglycerate dehydrogenase variants responsible for pathological reduced L‐serine levels

Posted on 31 August 2023 by

In the brain, L-serine is produced through the phosphorylated pathway (PP). hPHGDH catalyzes the first and rate-limiting step in the PP. Three variants related to hPHGDH deficiency and Neu-Laxova syndrome have been studied. V261M, V425M, and V490M substitutions alter the kinetic and structural properties of hPHGDH. Variants ectopic expression results in protein aggregation and reduced L-serine level.

Abstract

In the brain, the non-essential amino acid L-serine is produced through the phosphorylated pathway (PP) starting from the glycolytic intermediate 3-phosphoglycerate: among the different roles played by this amino acid, it can be converted into D-serine and glycine, the two main co-agonists of NMDA receptors. In humans, the enzymes of the PP, namely phosphoglycerate dehydrogenase (hPHGDH, which catalyzes the first and rate-limiting step of this pathway), 3-phosphoserine aminotransferase, and 3-phosphoserine phosphatase are likely organized in the cytosol as a metabolic assembly (a “serinosome”). The hPHGDH deficiency is a pathological condition biochemically characterized by reduced levels of L-serine in plasma and cerebrospinal fluid and clinically identified by severe neurological impairment. Here, three single-point variants responsible for hPHGDH deficiency and Neu-Laxova syndrome have been studied. Their biochemical characterization shows that V261M, V425M, and V490M substitutions alter either the kinetic (both maximal activity and K _m for 3-phosphoglycerate in the physiological direction) and the structural properties (secondary, tertiary, and quaternary structure, favoring aggregation) of hPHGDH. All the three variants have been successfully ectopically expressed in U251 cells, thus the pathological effect is not due to hindered expression level. At the cellular level, mistargeting and aggregation phenomena have been observed in cells transiently expressing the pathological protein variants, as well as a reduced L-serine cellular level. Previous studies demonstrated that the pharmacological supplementation of L-serine in hPHGDH deficiencies could ameliorate some of the related symptoms: our results now suggest the use of additional and alternative therapeutic approaches.

Downregulation of microRNA‐326 enhances ZNF322A expression, transcriptional activity and tumorigenic effects in lung cancer

Posted on 30 August 2023 by

Oncogenic ZNF322A transcription factor is overexpressed in lung cancer. Downregulated miR-326 promotes ZNF322A-induced tumor growth and metastasis. This study reveals that miR-326-low/ZN322A-high profile is a biomarker to predict poor prognosis in lung cancer.

Abstract

Zinc finger protein ZNF322A is an oncogenic transcription factor. Overexpression of ZNF322A activates pro-metastasis, cancer stemness, and neo-angiogenesis-related genes to enhance lung cancer progression. However, the upstream regulator of ZNF322A is not well defined. Dysregulation of microRNAs (miRNAs) can mediate cancer cell growth, migration, and invasion to promote tumorigenesis. Here, we uncover the mechanism of miRNA-mediated transcriptional regulation in ZNF322A-driven oncogenic events. ZNF322A harbors several putative miRNA-binding sites in the 3′-untranslated region (UTR). We validated that miR-326 downregulated ZNF322A-3′-UTR luciferase activity and mRNA expression. Furthermore, miR-326 suppressed the expression of ZNF322A-driven cancer-associated genes such as cyclin D1 and alpha-adducin. Reconstitution experiments by ectopic overexpression of ZNF322A abolished miR-326-suppressed cancer cell proliferation and cell migration capacity. Moreover, miR-326 attenuated ZNF322A-induced tumor growth and lung tumor metastasis in vivo. Clinically, the expression of miR-326 negatively correlated with ZNF322A mRNA expression in surgically resected tissues from 120 non-small cell lung cancer (NSCLC) patients. Multivariate Cox regression analysis demonstrated that NSCLC patients with low miR-326/high ZNF322A profile showed poor overall survival. Our results reveal that the deregulated expression of miR-326 leads to hyperactivation of ZNF322A-driven oncogenic signaling. Targeting the miR-326/ZNF322A axis would provide new therapeutic strategies for lung cancer patients.

Advances in screening, synthesis, modification, and biomedical applications of peptides and peptide aptamers

Posted on 30 August 2023 by

This comprehensive review explores the latest advancements in the screening, synthesis, modification, and biomedical applications of peptides and peptide aptamers. It discusses screening techniques, diverse synthesis strategies, and various modification approaches employed to enhance their properties. The review also highlights the broad range of biomedical applications where peptides and peptide aptamers have shown promise, including drug delivery, therapeutics, diagnostics, and biomaterials.

Abstract

Peptides and peptide aptamers have emerged as promising molecules for a wide range of biomedical applications due to their unique properties and versatile functionalities. The screening strategies for identifying peptides and peptide aptamers with desired properties are discussed, including high-throughput screening, display screening technology, and in silico design approaches. The synthesis methods for the efficient production of peptides and peptide aptamers, such as solid-phase peptide synthesis and biosynthesis technology, are described, along with their advantages and limitations. Moreover, various modification techniques are explored to enhance the stability, specificity, and pharmacokinetic properties of peptides and peptide aptamers. This includes chemical modifications, enzymatic modifications, biomodifications, genetic engineering modifications, and physical modifications. Furthermore, the review highlights the diverse biomedical applications of peptides and peptide aptamers, including targeted drug delivery, diagnostics, and therapeutic. This review provides valuable insights into the advancements in screening, synthesis, modification, and biomedical applications of peptides and peptide aptamers. A comprehensive understanding of these aspects will aid researchers in the development of novel peptide-based therapeutics and diagnostic tools for various biomedical challenges.

Reduced iron and cobalt levels in response to curcumin supplementation are not responsible for the prolonged larval development and do not affect the oxidative stress tolerance and polyamine status of D. melanogaster

Posted on 19 August 2023 by

Curcumin supplementation, similar to the iron chelator bathophenanthroline, lowered the iron but also the cobalt content, and to a lesser extent affected the manganese and zinc status of D. melanogaster. However, these alterations in trace metal balance did not affect catalase activity, oxidative stress tolerance, and polyamine status in fruit flies. In contrast, in curcumin-supplemented mice, the spleen exhibits an elevated spermidine production, which is most probably related to a compensatory growth due to curcumin-induced iron deficiency.

Abstract

Recent reports indicated that the phytochemical curcumin possesses iron-chelating activity. Here, by employing the fruit fly Drosophila melanogaster, we conducted feeding studies supplementing curcumin or, as a control, the iron chelator bathophenanthroline (BPA). First, the absorption and further metabolization of dietary curcuminoids were proved by metabolomics analyses. Next, we found that 0.2% dietary curcumin, similar to BPA, lowered the iron but also the cobalt content, and to a lesser extent affected the manganese and zinc status. Supplementation during larval stages was required and sufficient for both compounds to elicit these alterations in adult animals. However, curcumin-induced retarded larval development was not attributable to the changed trace metal status. In addition, a reduction in the iron content of up to 70% by curcumin or BPA supplementation did not reduce heme-dependent catalase activity and tolerance toward H₂O₂ in D. melanogaster. Moreover, polyamines were not influenced by curcumin treatment and decreased iron levels. This was confirmed for selected organs from 0.2% curcumin-treated mice, except for the spleen. Here, elevated spermidine level and concomitant upregulation of genes involved in polyamine production were associated with a putatively anemia-derived increased spleen mass. Our data underline that the metal-chelating property of curcumin needs to be considered in feeding studies.

Dapagliflozin improves diabetic cognitive impairment via indirectly modulating the mitochondria homeostasis of hippocampus in diabetic mice

Posted on 19 August 2023 by

The goal of this study is to determine the underlying mechanism and look into dapagliflozin's impact on diabetic cognitive impairment. Diabetes-induced cognitive dysfunction was attenuated by dapagliflozin and the effect was indirect rather than direct.

Abstract

Cognitive impairment is increasingly recognized as an important comorbidity of diabetes progression; however, the underlying molecular mechanism is unclear. Dapagliflozin, an inhibitor of sodium-glucose co-transporter 2 (SGLT2), has shown promising effects against diabetes in rodent experiments and human clinical assays. This study aimed to determine the underlying mechanism and examine the effect of dapagliflozin on diabetic cognitive impairment. To create an in vivo model of diabetic cognitive impairment, streptozotocin (STZ)-induced diabetic mice were used. Dapagliflozin was administered to mice for 8 weeks. The context fear condition and Morris water maze test was used to evaluate mice's behavioral change. Western blotting was used to evaluate protein expression. Hematoxylin and eosin (HE) and Nissl staining were applied to monitor morphological and structural changes. Congo red staining was performed to identify the formation of senile plaques. Mitochondria morphology was examined using a transmission electron microscope, and blood flow in the mouse cerebral cortex was measured using a laser Doppler imaging assay. Comparison to the diabetes mellitus (DM) group, the dapagliflozin group had lower glucose levels. Behavioral studies have shown that dapagliflozin can restore memory deficits in diabetic mice. The murky cell membrane edges and Nissl bodies more difficult to identify in the DM group were revealed by HE and Nissl staining, which were both improved by dapagliflozin treatment. Dapagliflozin inhibited the progression of Aβ generation and the reduced cerebral blood flow in the DM group was rescued. After dapagliflozin treatment, damaged mitochondria and lack of SGLT2 in the hippocampus and cortex of diabetic mice were repaired. Diabetes-induced cognitive dysfunction was attenuated by dapagliflozin and the effect was indirect rather than direct.

Bioactive peptides in preterm human milk: Impact of maternal characteristics and their association to neonatal outcomes

Posted on 16 August 2023 by

Hormone content of very preterm human milk is partially determined by maternal factors most of them regarding metabolic health and obstetric characteristics. Human milk concentration impacts on growth and development of preterm infants during their stay in the neonatal unit.

Abstract

Human milk adipokines in term babies seem partially determined by maternal factors and affect infant's development. We aimed to describe bioactive peptide concentration in very preterm human milk and associations to maternal characteristics and postnatal growth. Mothers delivering ≤32 weeks of gestation and their infant/s were recruited. At 4 weeks of lactation, an aliquot of 24-h-pooled milk was collected for exclusively breastfeeding dyads. Insulin, leptin, adiponectin, and milk fat globule epidermal growth factor-8 (MFG-E8) were measured by enzyme-linked immunoabsorbent assay in skimmed milk. One hundred mothers (28.8 ± 2.3 weeks at delivery) provided a milk sample. Milk insulin was related to gestational age, pre-pregnancy body mass index (BMI), and galactagogue treatment (final model: adjusted R ²: 0.330, p < 0.0001; adjusted β coefficients: galactagogue treatment: 0.348, p 0.001; pre-pregnancy BMI: 0.274, p 0.009; gestational age: −0.290, p 0.007). Adiponectin was higher in mothers with gestational diabetes (30.7 ± 6.5 vs. 24.8 ± 8 ng/mL, p 0.044). Leptin was associated with pre-pregnancy BMI (Spearman's ρ: 0.648, p < 0.0001) and MFG-E8 to presence of labor and multiple pregnancy (final linear regression model, R ²: 0.073, p 0.028, adjusted β coefficients: presence of labor −0.229, p 0.050; twins: −0.192, p 0.099). Milk adiponectin was associated with a greater decrease in length z-scores from birth to 28 days (Pearson's r: −0.225, p 0.032) and to discharge (Pearson's r: −0.290, p 0.003). Milk MFG-E8 was lower in milk of mothers whose babies experienced late-onset sepsis (13.3 ± 5.8 vs. 16.8 ± 6.3 μg/mL, p 0.023). Adipokines levels in preterm human milk are partially related to maternal metabolic status. Milk peptide concentration associates with early neonatal growth trajectories.

The role of P450 enzymes in malaria and other vector‐borne infectious diseases

Posted on 9 August 2023 by nOleksii Skorokhod, nEkaterina Vostokova, nGianfranco Gilardin

We review published research data on P450 enzymes from all players in vector-borne infections, that is, pathogens, vectors, and hosts, regarding the potential role of CYPs in disease. We discuss strategies on how to exploit cytochromes P450 in vector-borne disease control.

Abstract

Vector-borne infectious diseases are still an important global health problem. Malaria is the most important among them, mainly pediatric, life-threatening disease. Malaria and other vector-borne disorders caused by parasites, bacteria, and viruses have a strong impact on public health and significant economic costs. Most vector-borne diseases could be prevented by vector control, with attention to the ecological and biodiversity conservation aspects. Chemical control with pesticides and insecticides is widely used as a measure of prevention although increasing resistance to insecticides is a serious issue in vector control. Metabolic resistance is the most common mechanism and poses a big challenge. Insect enzyme systems, including monooxygenase CYP P450 enzymes, are employed by vectors mainly to metabolize insecticides thus causing resistance. The discovery and application of natural specific inhibitors/blockers of vector P450 enzymes as synergists for commonly used pesticides will contribute to the “greening” of insecticides. Besides vector CYPs, host CYP enzymes could also be exploited to fight against vector-borne diseases: using mostly their detoxifying properties and involvement in the immune response. Here, we review published research data on P450 enzymes from all players in vector-borne infections, that is, pathogens, vectors, and hosts, regarding the potential role of CYPs in disease. We discuss strategies on how to exploit cytochromes P450 in vector-borne disease control.

Caffeic acid phenethyl ester surmounts acquired resistance of AZD9291 in non‐small cell lung cancer cells

Posted on 9 August 2023 by nYaw Twum, nKent Marshall, nWeimin Gaon

We developed and characterized a new tyrosine kinase inhibitor resistant non-small cell lung cancer (NSCLC) line, HCC827GR, which (1) was significantly resistant to both gefitinib and AZD9291; (2) had more in vitro tumorigenic potential as evident by a larger colony size from the anchorage-independent growth assay; and (3) exhibited a total of 26 differentially expressed genes (≥two fold-change) as compared to HCC827. These genes were involved in regulating cell growth, transcription, phase 1 metabolism, cell cycle, and apoptosis. Additionally, AZD9291 in combination with CAPE partially reverted the AZD9291 resistance in HCC827GR cells. This was exhibited by (1) synergistically suppressed cell viability; (2) cell cycle arrest and apoptosis induced through suppressing EGFR activation and modulating p53, p21, cyclin D1, and survivin protein expressions; and (3) differentially regulating genes involved in cancer drug resistance pathways.

Abstract

Epithelial growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are the first-line therapy for EGFR mutated non-small cell lung cancer (NSCLC); however, resistance rapidly develops. The objective of this study was therefore to establish and characterize a gefitinib resistant NSCLC line (HCC827GR) and evaluate the therapeutic effects of natural products in combination with third-generation EGFR-TKI, AZD9291. The IC₅₀ of gefitinib and AZD9291 in HCC827GR were significantly higher than those of HCC827 (p < 0.05). Furthermore, anchorage-independent colony assay indicated that HCC827GR cells were more aggressive than their predecessors. This was reflected by the gene/protein expression changes observed in HCC827GR versus HCC827 profiled by cancer drug resistance real-time polymerase chain reaction (RT-PCR) array and Western blot. Three natural products were screened and caffeic acid phenethyl ester (CAPE) exhibited the most significant combinative cytotoxic effect with AZD9291. Specifically, flow cytometry revealed that AZD9291 + CAPE considerably increased the fraction of cell in pre-G1 of the cell cycle and caspase-Glo3/7 assay showed a dramatic increase in apoptosis when compared to AZD9291 alone. Furthermore, Western blot showed significant downregulation of p-EGFR/p-AKT in HCC827GR cells treated with AZD9291 + CAPE as compared to AZD9291. Moreover, it is evident that AZD9291 + CAPE specifically resulted in a marked reduction in the protein expressions of the cell-proliferation-related genes p21, cyclin D1, and survivin. Finally, refined RT-PCR/Western blot data indicated that AZD9291 + CAPE may at least partially exert its synergistic effects via the PLK2 pathway. Together, these results suggest that CAPE is a clinically relevant compound to aid AZD9291 in treating EGFR-TKI resistant cells through modulating critical genes/proteins involved in cancer resistance/therapy.

Roles of circular RNAs in osteogenic/osteoclastogenic differentiation

Posted on 3 August 2023 by nTao Wang, nChao Zhang, nLin Xu, nXingnuan Lin

The present review provides a systematic overview of recent literature on the processes through which circRNAs regulate the dynamic balance between osteoblasts and osteoclasts that ultimately preserve bone homeostasis. It will also give insight that can shape current understanding of the pathogenesis of OP and other bone metabolic diseases to better guide diagnostic and treatment strategies for affected patients.

Abstract

The process of bone remodeling occurs and is regulated through interactions between osteoclasts, which resorb bone, and osteoblasts, which generate bone tissue. When the homeostatic balance between these two cell types is dysregulated, this can contribute to abnormal bone remodeling resulting in a loss of bone mass as is observed in osteoporosis (OP) and other forms of degenerative bone metabolic diseases. At present, details of molecular mechanism underlying the development of bone metabolic diseases such as OP remain to be elucidated. Circular RNAs (circRNAs) are small non-coding RNA molecules with a closed-loop structure that can regulate the differentiation of osteoclasts and osteoblasts. The present review provides a systematic overview of recent literature on the processes through which circRNAs regulate the dynamic balance between osteoblasts and osteoclasts that ultimately preserve bone homeostasis. It will also give insight that can shape current understanding of the pathogenesis of OP and other bone metabolic diseases to better guide diagnostic and treatment strategies for affected patients.