Aberrant Cell Proliferation Research Articles

Parvimonas micra infection enhances proliferation, wound healing, and inflammation of a colorectal cancer cell line.

The gut microbiota Parvimonas micra has been found to be enriched in gut mucosal tissues and fecal samples of colorectal cancer (CRC) patients compared with non-CRC controls. In the present study, we investigated the tumorigenic potential of P. micra and its regulatory pathways in CRC using HT-29, a low-grade CRC intestinal epithelial cell. For every P. micra-HT-29 interaction assay, HT-29 was co-cultured anaerobically with P. micra at an MOI of 100:1 (bacteria: cells) for 2 h. We found that P. micra increased HT-29 cell proliferation by 38.45% (P=0.008), with the highest wound healing rate at 24 h post-infection (P=0.02). In addition, inflammatory marker expression (IL-5, IL-8, CCL20, and CSF2) was also significantly induced. Shotgun proteomics profiling analysis revealed that P. micra affects the protein expression of HT-29 (157 up-regulated and 214 down-regulated proteins). Up-regulation of PSMB4 protein and its neighbouring subunits revealed association of the ubiquitin-proteasome pathway (UPP) in CRC carcinogenesis; whereas down-regulation of CUL1, YWHAH, and MCM3 signified cell cycle dysregulation. Moreover, 22 clinically relevant epithelial-mesenchymal transition (EMT)-markers were expressed in HT-29 infected with P. micra. Overall, the present study elucidated exacerbated oncogenic properties of P. micra in HT-29 via aberrant cell proliferation, enhanced wound healing, inflammation, up-regulation of UPPs, and activation of EMT pathways.

FAR-INFRARED IRRADIATION INHIBITS CELL PROLIFERATION IN BASAL AND PDGF-STIMULATED RAT VASCULAR SMOOTH MUSCLE CELLS THROUGH REGULATION AMPK/MTOR/P70S6K SIGNALING AXIS

Objective: Far-infrared (FIR) rays are invisible electromagnetic radiation with a wavelength of 3-1000 um, which elicits molecular vibrations in water and other organic molecules in cells and tissues, thereby FIR rays may have various biological effects. Aberrant proliferation of vascular smooth muscle cells (VSMCs) is associated with the development of diverse vascular diseases including atherosclerosis. Although FIR irradiation is reported to inhibit cell proliferation in various types of cells, effect of FIR irradiation on VSMC proliferation and the underlying mechanism remain elusive. Here, we investigated the molecular mechanism by which FIR irradiation inhibited VSMC proliferation. Design and method: We performed cell proliferation and viability assay, inhibitor studies, transfection of dominant negative (dn)-AMP-activated protein kinase (AMPK) alpha 1 gene, and western blot analyses. We also conducted confocal microscopic image analyses and ex vivo studies using isolated rat aortas. Results: FIR irradiation for 30 min decreased VSMC proliferation with no alteration in cell viability, which was accompanied by decreased phosphorylation of mammalian target of rapamycin (mTOR) at Ser2448 (p-mTOR-Ser2448) and p70 S6 kinase (p70S6K) at Thr389 (p-p70S6K-Thr389). Furthermore, FIR irradiation increased phosphorylation of AMP-activated protein kinase (AMPK) at Thr172 (p-AMPK-Thr172). Pre-treatment with compound C, a specific AMPK inhibitor, or ectopic expression of dn-AMPK alpha 1 gene, significantly reversed FIR irradiation-decreased VSMC proliferation, p-mTOR-Ser2448, and p-p70S6K-Thr389. However, hyperthermal stimulus (39 degrees) did not change VSMC proliferation and AMPK/mTOR/p70S6K phosphorylation. Similar to in vitro results, FIR irradiation increased p-AMPK-Thr172, and decreased p-mTOR-Ser2448 and p-p70S6K-Thr389 in isolated rat aortas. Finally, FIR irradiation attenuated platelet-derived growth factor (PDGF)-stimulated VSMC proliferation by increasing p-AMPK-Thr172, and decreasing p-mTOR-Ser2448 and p-p70S6K-Thr389 in PDGF-induced in vitro atherosclerosis model. Conclusions: We demonstrate that FIR irradiation decreases VSMC proliferation, at least in part, by regulating AMPK/mTOR/p70S6K signaling axis. These observations suggest that FIR ray therapy can be used to treat arterial narrowing diseases including atherosclerosis and in-stent restenosis.

Development of bifunctional CRBN-SOS1 degraders for treatment of mutant KRAS cancers.

3151 Background: KRAS, a GTPase that regulates cellular growth, is the most frequently mutated oncogene in non-small cell lung, pancreatic and colorectal cancers. Hyperactivation of KRAS in mutant alleles result in upregulated signaling within the MAPK pathway leading to aberrant cell proliferation. In addition to the pan- and mutant-specific KRAS inhibitors, its upstream GEF activator, SOS1, has emerged as an attractive therapeutic target given its ability to regulate the active, GTP-loaded state of KRAS and mitigate MAPK pathway feedback. Methods: Utilizing our PRODEGY platform, we developed bifunctional CRBN-SOS1 degraders for the treatment of patients with KRAS-mutated cancers, independent of mutation type. Results: Our degraders display CRBN- and proteasome-mediated SOS1 degradation with DC50s < 15nM in high-throughput (HiBiT) and traditional Western Blot assays in multiple mutant KRAS cell lines. SOS1 degradation resulted in the reduction of active KRAS, HRAS and MRAS levels when assayed by RAF-RBD pulldowns in MIA PaCa-2 (KRAS G12C) cells. Other downstream signaling markers (pERK and pS6) were also decreased upon SOS1 degradation in KRAS G12C, G12V, G12S and G13D cell lines in 3D Western Blot assays. Notably, the observed pERK and pS6 inhibition was driven by SOS1 degradation and not inhibition as decreases in signaling was no longer observed when SOS1 degraders were outcompeted with CRBN competitors. To assess the antiproliferative effects of our SOS1 degraders, we developed 3D proliferation assays in various KRAS mutant (G12A, G12C, G12D, G12V, G12S, G13C and G13D) as well as KRAS amplified cell lines and observed IC50s ranging from 0.5 - 70nM. Treatment of CRBN KO and SOS1 KO H358 and MIA PaCa-2 cell lines with our SOS1 degraders resulted in 100- to 400-fold IC50 values, validating that the functional activity observed is dependent on its targets. Consistent with our in vitro data, treatment of SOS1 degraders in MIA PaCa-2 xenografts resulted in > 90% degradation and led to significant tumor growth inhibition. Given the diversity of mutations and tumors as well as acquired drug resistance to allele-specific KRAS inhibitors, we sought to determine if coupling SOS1 degradation with other EGFR-RAS-MAPK inhibitors could produce more potent responses to maximize patient benefit. Strong synergistic antiproliferative effects were observed when SOS1 degraders were co-treated with KRAS inhibitors AMG510 (G12C) and MRTX1133 (G12D) in respective KRAS-mutant cell lines as well as with MEK inhibition and EGFR inhibition. Lastly, combination of AMG510 or Trametinib with SOS1 degraders yielded strong synergy and a greater anti-tumor response in MIA PaCa-2 xenografts relative to the individual agents themselves. Conclusions: Together, these results highlight the potential of SOS1 degraders alone and in combination as therapeutic options for a variety of KRAS-driven cancers.

Analysis of prognostic biomarker models and immune microenvironment in acute myeloid leukemia by integrative bioinformatics.

Acute myeloid leukemia (AML) is a hematological cancer driven on by aberrant myeloid precursor cell proliferation and differentiation. A prognostic model was created in this study to direct therapeutic care. Differentially expressed genes (DEGs) were investigated using the RNA-seq data from the TCGA-LAML and GTEx. Weighted Gene Coexpression Network Analysis (WGCNA) examines the genes involved in cancer. Find the intersection genes and construct the PPI network to discover hub genes and remove prognosis-related genes. A nomogram was produced for predicting the prognosis of AML patients using the risk prognosis model that was constructed using COX and Lasso regression analysis. GO, KEGG, and ssGSEA analysis were used to look into its biological function. TIDE score predicts immunotherapy response. Differentially expressed gene analysis revealed 1004 genes, WGCNA analysis revealed 19,575 tumor-related genes, and 941 intersection genes in total. Twelve prognostic genes were found using the PPI network and prognostic analysis. To build a risk rating model, RPS3A and PSMA2 were examined using COX and Lasso regression analysis. The risk score was used to divide the patients into two groups, and Kaplan-Meier analysis indicated that the two groups had different overall survival rates. Univariate and multivariate COX studies demonstrated that risk score is an independent prognostic factor. According to the TIDE study, the immunotherapy response was better in the low-risk group than in the high-risk group. We eventually selected out two molecules to construct prediction models that might be used as biomarkers for predicting AML immunotherapy and prognosis.

Multi-omics data integration reveals the molecular network of dysregulation IQGAP2-mTOR promotes cell proliferation.

IQGAP2 as a tumor suppressor gene can influence cell proliferation in multiple tumor cell lines. However, the regulation network of cell proliferation resulting solely from the deficiency of IQGAP2 in cells was still unclear. Here, we integrated transcriptome, proteome, and phosphoproteome analyses to investigate the regulatory network of cell proliferation in IQGAP2 knockdown HaCaT and HEK293 cells. Our findings revealed that the dysregulation of the IQGAP2-mTOR molecular network led to increased cell proliferation. We demonstrated that IQGAP2 knockdown enhanced the phosphorylation levels of AKT and S6K, leading to increased cell proliferation. Additionally, we found that AKT and mTOR inhibitors partially rescued abnormal cell proliferation by reducing hyperphosphorylation. Our data suggest a potential connection between the mTOR signaling pathway and aberrant cell proliferation in IQGAP2 knockdown cells. These findings offer a new therapeutic strategy for patients with IQGAP2 deficiency.

Computational Exploration of Functional and Structural Impact of Single Nucleotide Changes in DNMT3A Gene among Acute Myeloid Leukemia Patients

Acute myeloid leukemia (AML) is a blood cell malignancy of the myeloid line, characterized by fast proliferation of aberrant cells that build up in the bone marrow and blood, interfering with normal blood cell synthesis. DNMT3A is a DNA methyltransferase that plays a role in DNA methylation, an epigenetic modification associated with gene expression regulation. DNMT3A mutations are frequently found in AML and are associated with poor prognosis. Objective: To evaluate the impact of DNMT3A mutations on protein structure and function, specifically in the context of AML. Methods: SNPs of DNMT3A gene reported in AML (R882P, R882L, R882S, R882G, and R882C) were retrieved from National Centre for Biotechnology Information (NCBI) database and different in silico approaches were used to investigate how these mutations affect protein structure and function. Results: Prediction tools indicated that mutations are pathogenic affecting DNMT3A function and were found in evolutionarily conserved regions. Protein stability analysis showed that mutations reduce DNMT3A's structural stability, alter secondary structure of the protein, particularly helices, interacts with other proteins and reduce protein-protein affinity. RNA folding analysis revealed abnormal folding patterns caused by mutant, affecting protein translation. DNMT3A expression was reported to be considerably greater in AML compared to normal tissues, and mutations were associated with poor overall survival in AML patients. Methylation levels and post-translational modification sites of DNMT3A were also investigated. Conclusions: Overall, this research highlighted the negative impact of DNMT3A mutations on protein structure and function, emphasizing their importance in the development and prognosis of AML. Acute myeloid leukemia (AML) is a blood cell malignancy of the myeloid line, characterized by fast proliferation of aberrant cells that build up in the bone marrow and blood, interfering with normal blood cell synthesis. DNMT3A is a DNA methyltransferase that plays a role in DNA methylation, an epigenetic modification associated with gene expression regulation. DNMT3A mutations are frequently found in AML and are associated with poor prognosis. Objective: To evaluate the impact of DNMT3A mutations on protein structure and function, specifically in the context of AML. Methods: SNPs of DNMT3A gene reported in AML (R882P, R882L, R882S, R882G, and R882C) were retrieved from National Centre for Biotechnology Information (NCBI) database and different in silico approaches were used to investigate how these mutations affect protein structure and function. Results: Prediction tools indicated that mutations are pathogenic affecting DNMT3A function and were found in evolutionarily conserved regions. Protein stability analysis showed that mutations reduce DNMT3A's structural stability, alter secondary structure of the protein, particularly helices, interacts with other proteins and reduce protein-protein affinity. RNA folding analysis revealed abnormal folding patterns caused by mutant, affecting protein translation. DNMT3A expression was reported to be considerably greater in AML compared to normal tissues, and mutations were associated with poor overall survival in AML patients. Methylation levels and post-translational modification sites of DNMT3A were also investigated. Conclusions: Overall, this research highlighted the negative impact of DNMT3A mutations on protein structure and function, emphasizing their importance in the development and prognosis of AML.

Homeobox A9 is a novel mediator of vascular smooth muscle cell phenotypic switching and proliferation by regulating methyl-CpG binding protein 2

Aberrant proliferation and phenotypic switching of vascular smooth muscle cells (VSMCs) are considered to be the main pathological processes of atherosclerotic plaque formation. Methyl-CpG binding protein 2 (MECP2) affects cell differentiation via modulating VSMC-specific gene expression and acts as a driver for the development of atherosclerosis (AS). Here, we aimed to elucidate the role of homeobox A9 (HOXA9) on aberrant VSMCs upon injury or AS, and whether HOXA9-mediated VSMC injury was associated with MECP2. Adeno-associated virus serotype 8-mediated knockdown of HOXA9 rescued aortic pathological injury of apolipoprotein E-deficient (ApoE−/−) mice fed a high-fat diet (HFD), characterized by the reduction of lipid accumulation and foam cell formation. Further in vitro evidence suggested that proliferation and migration of primary mouse VSMCs (mVSMCs) stimulated by oxidized low-density lipoprotein (ox-LDL) were inhibited after HOXA9 silencing. In addition, HOXA9 silencing blocked VSMC phenotypic switching from contractile to a pathological synthetic state. HOXA9 overexpression caused opposite alterations in ox-LDL-stimulated mVSMCs. Mechanistically, MECP2 was transcriptionally activated by HOXA9. Forced expression of MECP2 impaired the anti-proliferation, anti-migration, and phenotypic switching abilities of HOXA9 silencing in VSMCs upon ox-LDL stimulation. Collectively, our findings reveal that the role of HOXA9 in pathological vascular remodeling may attribute to transcriptional regulation of MECP2.

Mitochondrial complex III deficiency drives c-MYC overexpression and illicit cell cycle entry leading to senescence and segmental progeria

Accumulating evidence suggests mitochondria as key modulators of normal and premature aging, yet whether primary oxidative phosphorylation (OXPHOS) deficiency can cause progeroid disease remains unclear. Here, we show that mice with severe isolated respiratory complex III (CIII) deficiency display nuclear DNA damage, cell cycle arrest, aberrant mitoses, and cellular senescence in the affected organs such as liver and kidney, and a systemic phenotype resembling juvenile-onset progeroid syndromes. Mechanistically, CIII deficiency triggers presymptomatic cancer-like c-MYC upregulation followed by excessive anabolic metabolism and illicit cell proliferation against lack of energy and biosynthetic precursors. Transgenic alternative oxidase dampens mitochondrial integrated stress response and the c-MYC induction, suppresses the illicit proliferation, and prevents juvenile lethality despite that canonical OXPHOS-linked functions remain uncorrected. Inhibition of c-MYC with the dominant-negative Omomyc protein relieves the DNA damage in CIII-deficient hepatocytes in vivo. Our results connect primary OXPHOS deficiency to genomic instability and progeroid pathogenesis and suggest that targeting c-MYC and aberrant cell proliferation may be therapeutic in mitochondrial diseases.

Knockout of miR-184 in zebrafish leads to ocular abnormalities by elevating p21 levels.

miR-184 is one of the most abundant miRNAs expressed in the lens and corneal tissue. Mutations in the seed region of miR-184 are responsible for inherited anterior segment dysgenesis. Animal models recapitulating miR-184-related anterior segment dysgenesis are still lacking, and the molecular basis of ocular abnormalities caused by miR-184 dysfunction has not been well elucidated in vivo. In the present study, we constructed a miR-184-/- zebrafish line by destroying both two dre-mir-184 paralogs with CRISPR-Cas9 technology. Although there were no gross developmental defects, the miR-184-/- zebrafish displayed microphthalmia and cataract phenotypes. Cytoskeletal abnormalities, aggregation of γ-crystallin, and lens fibrosis were induced in miR-184-/- lenses. However, no obvious corneal abnormalities were observed in miR-184-/- zebrafish. Instead of apoptosis, deficiency of miR-184 led to aberrant cell proliferation and a robust increase in p21 levels in zebrafish eyes. Inhibition of p21 by UC2288 compromised the elevation of lens fibrosis markers in miR-184-/- lenses. RNA-seq demonstrated that levels of four transcriptional factors HSF4, Sox9a, CTCF, and Smad6a, all of which could suppress p21 expression, were reduced in miR-184-/- eyes. The predicted zebrafish miR-184 direct target genes (e.g., atp1a3a and nck2a) were identified and verified in miR-184-/- eye tissues. The miR-184-/- zebrafish is the first animal model mimicking miR-184-related anterior segment dysgenesis and could broaden our understanding of the roles of miR-184 in eye development.

Neurite outgrowth induced by stimulation of angiotensin II AT2 receptors in SH-SY5Y neuroblastoma cells involves c-Src activation

Neuroblastoma, the most common extracranial solid tumor occurring in childhood, originates from the aberrant proliferation of neural crest cells. Accordingly, the mechanism underling neuronal differentiation could provide new strategies for neuroblastoma treatment. It is well known that neurite outgrowth could be induced by Angiotensin II (Ang II) AT2 receptors; however, the signaling mechanism and its possible interaction with NGF (neural growth factor) receptors remain unclear. Here, we show that Ang II and CGP42112A (AT2 receptor agonist) promote neuronal differentiation by inducing neurite outgrowth and βIII-tubulin expression in SH-SY5Y neuroblastoma cells. In addition, we demonstrate that treatment with PD123319 (AT2 receptor antagonist) reverts Ang II or CGP42112A-induced differentiation. By using specific pharmacological inhibitors we established that neurite outgrowth induced by CGP42112A requires the activation of MEK (mitogen-activated protein kinase kinase), SphK (sphingosine kinase) and c-Src but not PI3K (phosphatidylinositol 3-kinase). Certainly, CGP42112A stimulated a rapid and transient (30 s, 1 min) phosphorylation of c-Src at residue Y416 (indicative of activation), following by a Src deactivation as indicated by phosphorylation of Y527. Moreover, inhibition of the NGF receptor tyrosine kinase A (TrkA) reduced neurite outgrowth induced by Ang II and CGP42112A. In summary, we demonstrated that AT2 receptor-stimulated neurite outgrowth in SH-SY5Y cells involves the induction of MEK, SphK and c-Src and suggests a possible transactivation of TrkA. In that regard, AT2 signaling pathway is a key player in neuronal differentiation and might be a potential target for therapeutic treatments.

Mitochondrial reactive oxygen species promote mitochondrial damage in high glucose-induced dysfunction and apoptosis of human dental pulp cells

Background/purposeHigh glucose (HG)-induced aberrant proliferation, apoptosis and odontoblastic differentiation of dental pulp cells (DPCs) have been implicated in the pathogenesis of impaired diabetic pulp healing; however, the underlying mechanism remains unclear. This study aimed to investigate the role of mitochondrial reactive oxygen species (mtROS) and mitochondria in HG-induced dysfunction and apoptosis of DPCs. Materials and methodsHuman DPCs (hDPCs) were cultured in a low-glucose, high-glucose, mannitol, and MitoTEMPO medium in vitro. Methylthiazol tetrazolium assay, Annexin V-FITC/PI staining and scratch-wound assay were used to analyze cell proliferation, apoptosis and migration, respectively. Alkaline phosphatase staining and alizarin red S staining were used to evaluate cell differentiation. DCF-DA staining, MitoSOX staining, MitoTracker Red staining, JC-1 staining, and adenosine triphosphate (ATP) kit assay were performed to investigate total ROS and mtROS generation, mitochondrial density, mitochondrial membrane potential (MMP), and ATP synthesis, respectively. Quantitative PCR assay was performed to detect the mRNA expression of mitochondrial biogenesis- and dynamics-related markers. Transmission electron microscopy was used to observe the mitochondrial ultrastructure. ResultsHG augmented the production of total ROS and mtROS, and triggered mitochondrial damage in hDPCs, as reflected by decreased mitochondrial density, depolarized MMP, reduced ATP synthesis, altered mRNA expression of mitochondrial biogenesis- and dynamics-related markers, and abnormal mitochondrial ultrastructure. Supplementation of MitoTEMPO alleviated the mitochondrial damage and reversed the aberrant proliferation, apoptosis, migration and odontoblastic differentiation of HG-stimulated hDPCs. ConclusionHG triggers mitochondrial damage via augmenting mtROS generation, resulting in the inhibited proliferation, migration, and odontoblastic differentiation of hDPCs and enhanced their apoptosis.

Principles, Processes and Modalities of Interaction Between Proto-Oncoprotein PIM-1 Kinase and Cell Division Cyclin CDC25A

All malignant tumors with aberrant cell differentiation and proliferation, unchecked growth, invasion, and metastasis are referred to as cancer. Its occurrence is a multi-step, complex process with many contributing components. Proto oncoprotein and The PIM kinase family is crucial to this procedure., and proto oncoprotein PIM-1 kinase is an important member of PIM family. The expression of proto oncoprotein PIM-1 is positively correlated with the status of lymphatic metastasis, and its overexpression could promote cell cycle and inhibit apoptosis. Cell division cycle 25 (CDC25) is a bispecific phosphatase from Schizosaccharomyces cerevisiae in 1986. CDC25 can activate CDK by catalyzing the dephosphorylation of inhibitory phosphorylation sites in the cell cycle dependent protein kinases (CDK) molecule, and regulate the response to DNA damage, thus promoting cell cycle operation. The interaction between the proto oncoprotein CDC25A in the above process is one of the keys to the current mechanism research with PIM-1 kinase. In this paper, PIM-1 and CDC25 are first introduced, respectively. Then, the interaction between PIM-1 and different substrates with a focus on CDC25A is presented. Finally, their role in tumor initiation and the application of CDC25A as a therapeutic target are highlighted. Therefore, this paper will discuss the interaction between CDC25A and PIM family.

Therapeutic Targets in Myelodysplastic Neoplasms: Beyond Hypomethylating Agents.

To discuss novel targeted therapies under investigation for treatment of myelodysplastic neoplasms (MDS). Over the last few years, results of phase 3 trials assessing novel therapies for high-risk MDS have been largely disappointing. Pevonedistat (NEDD-8 inhibitor) and APR-246 (TP53 reactivator) both did not meet trial endpoints. However, early phase trials of BCL-2, TIM3, and CD47 inhibitors have shown exciting data and are currently under phase 3 investigation. Moreover, combination of hypomethylating agents (HMA) with novel therapies targeting the mutational (IDH, FLT3, spliceosome complex) or immune (PD-1/PDL-1, TIM-3, IRAK-4) pathways are being investigated in early phase clinical trials and have shown adequate safety and promising efficacy. Myelodysplastic neoplasms (MDS) are a group of hematopoietic neoplasms defined by cytopenias and morphological dysplasia. They are characterized by clonal proliferation of aberrant hematopoietic stem cells caused by recurrent genetic abnormalities. This leads to ineffective erythropoiesis, peripheral blood cytopenias, abnormal cell maturation, and a high risk of transformation into acute myeloid leukemia (AML). Allogeneic hematopoietic stem cell transplantation is the only curative therapy; however, it is not a suitable option for majority patients due to their age, comorbidities, and the high rate of treatment-related complications. HMAs remain the only FDA-approved treatment option for high-risk MDS. Due to intolerance, primary, and secondary resistance to HMA, there is a large unmet need to develop new safe and effective therapies for patients with MDS. In this review, we focus on the current management strategies and novel therapies in development for treatment of high-risk MDS.

Enhanced Lyn Activity Causes Severe, Progressive Emphysema and Lung Cancer.

The epidemiological patterns of incident chronic obstructive pulmonary disease (COPD) and lung adenocarcinoma are changing, with an increasing fraction of disease occurring in patients who are never-smokers or were not exposed to traditional risk factors. However, causative mechanism(s) are obscure. Overactivity of Src family kinases (SFKs) and myeloid cell-dependent inflammatory lung epithelial and endothelial damage are independent candidate mechanisms, but their pathogenic convergence has not been demonstrated. Here we present a novel preclinical model in which an activating mutation in Lyn, a nonreceptor SFK that is expressed in immune cells, epithelium, and endothelium-all strongly implicated in the pathogenesis of COPD-causes spontaneous inflammation, early-onset progressive emphysema, and lung adenocarcinoma. Surprisingly, even though activated macrophages, elastolytic enzymes, and proinflammatory cytokines were prominent, bone marrow chimeras formally demonstrated that myeloid cells were not disease initiators. Rather, lung disease arose from aberrant epithelial cell proliferation and differentiation, microvascular lesions within an activated endothelial microcirculation, and amplified EGFR (epidermal growth factor receptor) expression. In human bioinformatics analyses, LYN expression was increased in patients with COPD and was correlated with increased EGFR expression, a known lung oncogenic pathway, and LYN was linked to COPD. Our study shows that a singular molecular defect causes a spontaneous COPD-like immunopathology and lung adenocarcinoma. Furthermore, we identify Lyn and, by implication, its associated signaling pathways as new therapeutic targets for COPD and cancer. Moreover, our work may inform the development of molecular risk screening and intervention methods for disease susceptibility, progression, and prevention of these increasingly prevalent conditions.

Abstract 3415: Preclinical characterization of a brain penetrant RAF inhibitor, BDTX-4933, targeting oncogenic BRAF Class I/II/III and RAS mutations

Abstract Mutations in BRAF and RAS are often oncogenic and lead to a constitutively active MAPK pathway that promotes aberrant cell proliferation and tumor growth. Currently approved BRAF inhibitors are selective against monomeric BRAF V600 mutants. These drugs are largely inactive against non-V600 dimeric BRAF mutants and have poor brain penetration. Although there is an FDA-approved KRAS G12C mutant-selective inhibitor, there are no approved inhibitors for cancer patients who harbor other (non-G12C) KRAS and NRAS mutations which promote tumor growth likely through constitutively active RAF dimers. There remains a high unmet clinical need for a highly CNS penetrant oral RAF inhibitor that targets a broad spectrum of BRAF mutations and constitutively active RAF dimers without paradoxical activation of the MAPK signaling pathway. BDTX-4933 is a potent, reversible, CNS penetrant RAF MasterKey inhibitor designed to target a large family of oncogenic BRAF mutations including BRAF monomers and RAF dimers. The compound inhibits not only all classes (I, II, and III) of BRAF mutations but also targets constitutively active RAF dimers promoted by upstream oncogenic MAPK pathway alterations, such as RAS mutations. In a panel of cancer cell lines that endogenously express BRAF or RAS mutations, BDTX-4933 demonstrates inhibition of the MAPK pathway signaling without paradoxical activation, resulting in potent inhibition of cellular proliferation. BDTX-4933 shows target engagement, inhibiting ERK phosphorylation, in tumor models in vivo, achieving strong anti-tumor efficacy and tumor regression across tumor models driven by either BRAF or RAS mutations. Furthermore, BDTX-4933 exhibits high CNS exposure leading to dose-dependent tumor growth inhibition, and survival benefit in mice implanted intracranially with xenograft BRAF mutant tumors. BDTX-4933 has a best-in-class profile to treat cancer patients harboring BRAF mutations or RAF dimer-promoting upstream genetic alterations. IND-enabling studies for BDTX-4933 are on-going. Citation Format: Yoon-Chi Han, Pui-Yee Ng, Luisa Shin Ogawa, Shao Ning Yang, Miao Chen, Noboru Ishiyama, Tai-An Lin, Elizabeth Buck. Preclinical characterization of a brain penetrant RAF inhibitor, BDTX-4933, targeting oncogenic BRAF Class I/II/III and RAS mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3415.

A Deep Analysis of Brain Tumor Detection from MR Images Using Deep Learning Networks

Creating machines that behave and work in a way similar to humans is the objective of artificial intelligence (AI). In addition to pattern recognition, planning, and problem-solving, computer activities with artificial intelligence include other activities. A group of algorithms called “deep learning” is used in machine learning. With the aid of magnetic resonance imaging (MRI), deep learning is utilized to create models for the detection and categorization of brain tumors. This allows for the quick and simple identification of brain tumors. Brain disorders are mostly the result of aberrant brain cell proliferation, which can harm the structure of the brain and ultimately result in malignant brain cancer. The early identification of brain tumors and the subsequent appropriate treatment may lower the death rate. In this study, we suggest a convolutional neural network (CNN) architecture for the efficient identification of brain tumors using MR images. This paper also discusses various models such as ResNet-50, VGG16, and Inception V3 and conducts a comparison between the proposed architecture and these models. To analyze the performance of the models, we considered different metrics such as the accuracy, recall, loss, and area under the curve (AUC). As a result of analyzing different models with our proposed model using these metrics, we concluded that the proposed model performed better than the others. Using a dataset of 3264 MR images, we found that the CNN model had an accuracy of 93.3%, an AUC of 98.43%, a recall of 91.19%, and a loss of 0.25. We may infer that the proposed model is reliable for the early detection of a variety of brain tumors after comparing it to the other models.

Recent Perspective of Lactobacillus in Reducing Oxidative Stress to Prevent Disease.

During oxidative stress, an important factor in the development of many diseases, cellular oxidative and antioxidant activities are imbalanced due to various internal and external factors such as inflammation or diet. The administration of probiotic Lactobacillus strains has been shown to confer a range of antibacterial, anti-inflammatory, antioxidant, and immunomodulatory effects in the host. This review focuses on the potential role of oxidative stress in inflammatory bowel diseases (IBD), cancer, and liver-related diseases in the context of preventive and therapeutic effects associated with Lactobacillus. This article reviews studies in cell lines and animal models as well as some clinical population reports that suggest that Lactobacillus could alleviate basic symptoms and related abnormal indicators of IBD, cancers, and liver damage, and covers evidence supporting a role for the Nrf2, NF-κB, and MAPK signaling pathways in the effects of Lactobacillus in alleviating inflammation, oxidative stress, aberrant cell proliferation, and apoptosis. This review also discusses the unmet needs and future directions in probiotic Lactobacillus research including more extensive mechanistic analyses and more clinical trials for Lactobacillus-based treatments.

Inhibition of heterogeneous nuclear ribonucleoproteins A1 and oxidative stress reduces glycolysis via pyruvate kinase M2 in chronic thromboembolic pulmonary hypertension.

Chronic thromboembolic pulmonary hypertension (CTEPH) is a lethal complication of pulmonary embolism involving pulmonary artery occlusion and microvascular disease. The glucose metabolism and reactive oxygen species (ROS) production may be perturbed in CTEPH, but the precise mechanisms are unclear. This study investigated glucose metabolism in CTEPH employing pulmonary endarterectomy (PEA)-derived pulmonary artery smooth muscle cells (PASMCs) and characterized the roles of pyruvate kinase M2 (PKM2) and its regulation by heterogeneous nuclear ribonucleoproteins A1 (hnRNPA1) and ROS in CTEPH. PEA tissues and blood samples of CTEPH patients were collected to study the levels of PKM2. Primary PASMCs were isolated from PEA tissues. We used small interfering RNAs to knock down PKM2 and hnRNPAI, and applied antioxidant N-acetylcysteine (NAC) and mito-TEMPO to reduce ROS production. The expression of glucometabolic genes, ROS production, glycolysis rate and proliferative and migratory activities were analyzed in PEA-derived PASMCs. PKM2 levels in serum and PEA tissues of CTEPH patients were higher than that of the healthy controls. Compared to the control PASMCs, PEA-derived PASMCs showed increased PKM2 expression and ROS production. The rates of glycolysis, proliferation and migration were increased in PEA-PASMCs and could be mitigated by PKM2 downregulation through hnRNPA1 or ROS inhibition. Increased glycolysis and PKM2 expression were found in PEA-PASMCs. Inhibition of hnRNPA1 or ROS corrected the aberrant glycolysis, cell proliferation and migration by downregulating PKM2. Regulation of the hnRNPA1/PKM2 axis represents a potential therapeutic target for the treatment of CTEPH.

Experimental models of autism spectrum disorders on the example of the use of brain organelles

IntroductionASD are heterogeneous pathological conditions characterized by difficulties in establishing social contacts and the manifestation of repetitive behavior. An atypical trajectory of brain maturation, impaired neurogenesis, synaptogenesis, and an imbalance in the excitatory and inhibitory systems of the CNS form the morphofunctional basis of the ASD.ObjectivesTo understand the functioning of this complexly organized system in time and space, a three-dimensional model is needed. The closest in vitro model of the human brain from early embryonic stages to aging is brain organoids. Human brain organoids are self-organizing three-dimensional cell aggregates derived from pluripotent stem cells (hiPSCs)MethodsOrganelles generalize neurogenesis, gliogenesis, synaptogenesis, cell migration and cell differentiation, gyrification of the cerebral cortex, and reflect the connections of brain regions.ResultsThe use of telencephalon organelles in the RAS model revealed a deficit in neuronal migration, acceleration and disruption of cell cycle synchronization, aberrant cell proliferation, abundant synaptogenesis, temporary deviations in the development of the cortex, increased branching of neurons, unbalanced inhibitory differentiation of neurons, high activity of ion channels is a consequence of a violation of FOXG1 activity. Organelles generalize neurogenesis, gliogenesis, synaptogenesis, cell migration and cell differentiation, gyrification of the cerebral cortex, and reflect the connections of brain regions.The use of telencephalon organelles in the RAS model revealed a deficit in neuronal migration, acceleration and disruption of cell cycle synchronization, aberrant cell proliferation, abundant synaptogenesis, temporary deviations in the development of the cortex, increased branching of neurons, unbalanced inhibitory differentiation of neurons, high activity of ion channels is a consequence of a violation of FOXG1 activity .ConclusionshiPSCs can provide insight into the cellular mechanisms underlying ASD as a neuropsychiatric disorder, providing access to the development of platforms for in vitro drug screening and individualized patient therapy.Disclosure of InterestNone Declared

The Anti-Inflammatory Effects and Clinical Potential of Dexmedetomidine in Pulmonary Arterial Hypertension

A pathogenic aspect of pulmonary arterial hypertension (PAH) is the aberrant pulmonary arterial smooth muscle cell (PASMC) proliferation. PASMC proliferation is significantly affected by inflammation. A selective α-2 adrenergic receptor agonist called dexmedetomidine (DEX) modulates specific inflammatory reactions. We investigated the hypothesis that anti-inflammatory characteristics of DEX could lessen PAH that monocrotaline (MCT) causes in rats. In vivo, male Sprague-Dawley rats aged 6 weeks were subcutaneously injected with MCT at a dose of 60 mg/kg. Continuous infusions of DEX (2 µg/kg per hour) were started via osmotic pumps in one group (MCT plus DEX group) at day 14 following MCT injection but not in another group (MCT group). Right ventricular systolic pressure (RVSP), right ventricular end-diastolic pressure (RVEDP), and survival rate significantly improved in the MCT plus DEX group compared with the MCT group [RVSP, 34 mmHg ± 4 mmHg versus 70 mmHg ± 10 mmHg; RVEDP, 2.6 mmHg ± 0.1 mmHg versus 4.3 mmHg ± 0.6 mmHg; survival rate, 42% versus 0% at day 29 (P < 0.01)]. In the histologic study, the MCT plus DEX group showed fewer phosphorylated p65-positive PASMCs and less medial hypertrophy of the pulmonary arterioles. In vitro, DEX dose-dependently inhibited human PASMC proliferation. Furthermore, DEX decreased the expression of interleukin-6 mRNA in human PASMCs treated with fibroblast growth factor 2 (FGF2). These consequences suggest that DEX improves PAH by inhibiting PASMC proliferation through its anti-inflammatory properties. Additionally, DEX may exert anti-inflammatory effects via blocking FGF2-induced nuclear factor κ B activation. SIGNIFICANCE STATEMENT: Dexmedetomidine, a selective α-2 adrenergic receptor agonist utilized as a sedative in the clinical setting, improves pulmonary arterial hypertension (PAH) by inhibiting pulmonary arterial smooth muscle cell proliferation through its anti-inflammatory effect. Dexmedetomidine may be a new PAH therapeutic agent with vascular reverse remodeling effect.