High Proliferative Activity Research Articles

Blood biomarkers and Ki-67 proliferation marker in breast cancer

Metastasis is the leading cause of death in patients with breast cancer (BC). It is known that the lesion of regional lymph nodes by tumor cells is more common in tumors with higher proliferative activity. Moreover, there is literature evidence on effects of cytokines and proteins upon the migration potential of the tumor. The aim of our work was to study the correlation between the concentrations of cytokines, proteins, and expression of Ki-67 proliferation marker in breast cancer with histology of non-specific invasive carcinoma.On the basis of pathological findings, 16 patients had metastases in regional lymph nodes (group I), and 18 patients had no detectable metastases (group II). Solid-phase enzyme immunoassay was used to determine concentrations of 14 cytokines in the supernatants of immunocompetent blood cells, i.e., IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1ra, TNFα, IFNγ, G-CSF, GM-CSF, VEGF and MCP-1, and concentrations of 6 proteins were determined in blood serum: estrogen and progesterone receptors, cadherin-E (CDH1), plasminogen activator type 1 (PAI-1), mucin 1 (MUC1), heat shock protein 90αA1 (HSP90αA1). Immunohistochemical study of Ki-67 expression was performed in paraffin sections of tumors using monoclonal antibodies.The study showed that Ki-67 expression in tumor tissues and blood concentrations of IL-6, IL-8, IL-1β and TNFα were higher in group I patients. On the contrary, blood concentrations of CDH1 and PAI-1 were higher in group II patients. It was found that Ki-67 showed both inverse correlations with CDH1 and PAI1, and direct correlations with IL-8 and TNFα. CDH1 had a direct correlation with PAI1, and inverse correlations with IL-6, IL-1β and TNFα. The studied cytokines showed direct correlations with each other. The analysis of ROC curves showed good quality and optimal values of the cut-off points for Ki-67 expression, cytokine and protein concentrations, thus allowing best prediction for detectable lymphatic metastasis.On the basis of these results, a quotient was proposed, which represents a ratio of CDH1 contents to the sum of IL-1β and TNFα concentrations in blood samples, which can help identification of the patients with breast cancer at risk for lymphatic metastasis.

Abstract 2774: Moesin-induced adriamycin resistance through IL-6/MSN/STAT3 axis and therapeutic efficacy of atovaquone in chemotherapy resistant triple-negative breast cancer

Abstract Introduction: Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer with a high frequency of chemotherapy resistance. Moesin (MSN) is a cytoskeleton linker discovered as a novel oncogene in our previous proteomics analysis. Here, we evaluated the molecular mechanisms of MSN and how it functions in response to chemotherapy, which remains unclear in TNBC. Method: The 178 in-house TNBC patient cohort and METABRIC data set were enrolled for clinicopathologic analysis. The multiple TNBC four-cell lines were used to establish MSN knock-down and over-expressed in vitro models using a lentiviral system. RNA sequencing was analyzed with MDA-MB-231 parent cell and MDA-MB-231/ADR12 (adriamycin resistance cell line). Invasion and migration assay was done. The drug response based on MSN gene expression was observed by cell viability assay, western blot, and AV/PI flow cytometry. Drug resistance concerning cancer stemness was confirmed with sphere formation and ALDH activity assay. Immunofluorescence and immunoprecipitation were performed. The combination index(CI), which is to define the synergism of combined treatment was calculated with the Compusyn program. Results: The cohort with METABRIC and our in-house clinical data showed concordant findings that overexpressed MSN mRNA and protein expressions have a poor prognosis in TNBC patients. The MSN protein expression in TNBC was increased compared to the protein level in both the normal human tissues and non-TNBC cell lines. Overexpressed MSN induced high cell proliferation activity, invasion, and migration ability. The apoptosis rate (AV+/PI+) along with cleaved PARP and caspase-3 expression was increased in TNBC cell lines with MSN downregulation. Further tests to determine the role of MSN in adriamycin resistance showed an increased number and size of spheres in the MSN overexpressed cell lines. Moreover, the protein and mRNA levels of MSN expression in MDA-MB-231/ADR12 were significantly higher than the level in the matched parental cell line. MSN and phospho-STAT3(T705) were increased by treatment of IL-6 in TNBC cell lines. The direct binding between MSN and STAT3, and the co-localization of MSN and pSTAT3 were consistently confirmed by co-immunoprecipitation and immunofluorescence, which were all activated by IL-6. Blocking the STAT3 pathway with atovaquone restored ADR response in MSN overexpressed tumor cells. CI value on the combined treatment of ADR and atovaquone in the MDA-MB-231/ADR12 cell line showed higher synergetic efficacy than the parental cell line. Conclusion: This research demonstrates the involvement of the MSN in doxorubicin resistance through IL-6/MSN/STAT3 pathway and provides a novel therapeutic candidate for TNBC patients. Citation Format: Cheng Hyun Lee, Da Sol Kim, Soo Young Park, Han Suk Ryu. Moesin-induced adriamycin resistance through IL-6/MSN/STAT3 axis and therapeutic efficacy of atovaquone in chemotherapy resistant triple-negative breast cancer [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 2774.

Isolation and characterization of a novel hydrolase-producing probiotic Bacillus licheniformis and its application in the fermentation of soybean meal.

Soybean meal (SBM) is one of the most important sources of plant-based protein in the livestock and poultry industry. However, SBM contains anti-nutritional factors (ANFs) such as glycinin, β-conglycinin, trypsin inhibitor and phytic acid that can damage the intestinal health of animals, inevitably reducing growth performance. Fermentation using microorganisms with probiotic potential is a viable strategy to reduce ANFs and enhance the nutritional value of SBM. In this study, a novel potential probiotic Bacillus licheniformis (B4) with phytase, protease, cellulase and xylanase activity was isolated from camel feces. The ability of B4 to tolerate different pH, bile salts concentrations and temperatures were tested using metabolic activity assay. It was found that B4 can survive at pH 3.0, or 1.0% bile salts for 5 h, and displayed high proliferative activity when cultured at 50°C. Furthermore, B4 was capable of degrading glycinin, β-conglycinin and trypsin inhibitor which in turn resulted in significant increases of the degree of protein hydrolysis from 15.9% to 25.5% (p < 0.01) and crude protein from 44.8% to 54.3% (p < 0.001). After fermentation with B4 for 24 h, phytic acid in SBM was reduced by 73.3% (p < 0.001), the neutral detergent fiber (NDF) and the acid detergent fiber of the fermented SBM were significantly decreased by 38.40% (p < 0.001) and 30.20% (p < 0.05), compared to the unfermented SBM sample. Our results suggested that the effect of solid-state fermented SBM using this novel B. licheniformis (B4) strain, could significantly reduce phytic acid concentrations whilst improving the nutritional value of SBM, presenting itself as a promising alternative to phytase additives.

Oncolytic adenovirus-mediated dual knockdown of survivin and OCT4 improves therapeutic efficacy in esophageal cancer.

Survivin and octamer-binding transcription factor 4 (OCT4) are reportedly up-regulated in esophageal cancer (EC) and have been correlated with high tumor proliferative activity and poor prognosis. Oncolytic viruses encoding specific transgenes have been considered as therapeutic methods to increase therapeutic efficacy in a variety of solid tumors. In this study, an oncolytic adenovirus carrying short hairpin RNA (shRNA) of survivin (shSRVN) and OCT4 (shOCT4) was constructed to achieve dual knockdown of survivin and OCT4 and to explore the potential effect of the oncolytic adenovirus in EC. The oncolytic adenovirus replicated abundantly in human EC cells, with the replication multiplying by up to 192,085 and 620,055 times in esophageal carcinoma (Eca)-109 cells transfected with purified and completed recombinant adenoviruses called AdSProE1a-dual shRNA (shSRVN + shOCT4) and TE1 cells transfected with AdSProE1a-survivin shRNA (shSRVN) 96 hours after infection, respectively. The shRNAs targeting survivin and OCT4 significantly downregulated the expression levels of survivin and OCT4 in cells, thereby inhibiting the proliferative activity of cancer cells. Furthermore, E-cadherin and vimentin, which are both considered epithelial mesenchymal transition (EMT) markers, were found to be upregulated and downregulated, respectively, in cancer cells after exposure to the viral infection. The interference of survivin and OCT4 also contributed to cell cycle arrest and apoptosis, the half maximal inhibitory concentrations (IC50s) of oncolytic adenovirus loaded with AdSProE1a-shSRVN + shOCT4 in the Eca109 cells and the TE1 cells were 0.7271 and 0.1032 pfu/mL, respectively. Xenograft experiments in vivo showed that oncolytic adenovirus-mediated dual knockdown of survivin and OCT4 effectively inhibited the growth of xenografts and induced cancer cell apoptosis. We concluded that therapies targeting survivin and OCT4 have great potential for improving the therapeutic efficacy in EC. The dual target design strategy ensured the efficacy and safety of the treatment system and provided a novel and effective adjuvant target therapy for EC.

Slow progression of pediatric HIV associates with early CD8+ T cell PD-1 expression and a stem-like phenotype.

HIV nonprogression despite persistent viremia is rare among adults who are naive to antiretroviral therapy (ART) but relatively common among ART-naive children. Previous studies indicate that ART-naive pediatric slow progressors (PSPs) adopt immune evasion strategies similar to those described in natural hosts of SIV. However, the mechanisms underlying this immunophenotype are not well understood. In a cohort of early-treated infants who underwent analytical treatment interruption (ATI) after 12 months of ART, expression of PD-1 on CD8+ T cells immediately before ATI was the main predictor of slow progression during ATI. PD-1+CD8+ T cell frequency was also negatively correlated with CCR5 and HLA-DR expression on CD4+ T cells and predicted stronger HIV-specific T lymphocyte responses. In the CD8+ T cell compartment of PSPs, we identified an enrichment of stem-like TCF-1+PD-1+ memory cells, whereas pediatric progressors and viremic adults had a terminally exhausted PD-1+CD39+ population. TCF-1+PD-1+ expression on CD8+ T cells was associated with higher proliferative activity and stronger Gag-specific effector functionality. These data prompted the hypothesis that the proliferative burst potential of stem-like HIV-specific cytotoxic cells could be exploited in therapeutic strategies to boost the antiviral response and facilitate remission in infants who received early ART with a preserved and nonexhausted T cell compartment.

HBV-infected hepatocellular carcinoma can be robustly classified into three clinically relevant subgroups by a novel analytical protocol.

Liver cancer is the third leading cause of cancer-related death worldwide, and hepatocellular carcinoma (HCC) accounts for a relatively large proportion of all primary liver malignancies. Among the several known risk factors, hepatitis B virus (HBV) infection is one of the important causes of HCC. In this study, we demonstrated that the HBV-infected HCC patients could be robustly classified into three clinically relevant subgroups, i.e. Cluster1, Cluster2 and Cluster3, based on consistent differentially expressed mRNAs and proteins, which showed better generalization. The proposed three subgroups showed different molecular characteristics, immune microenvironment and prognostic survival characteristics. The Cluster1 subgroup had near-normal levels of metabolism-related proteins, low proliferation activity and good immune infiltration, which were associated with its good liver function, smaller tumor size, good prognosis, low alpha-fetoprotein (AFP) levels and lower clinical stage. In contrast, the Cluster3 subgroup had the lowest levels of metabolism-related proteins, which corresponded with its severe liver dysfunction. Also, high proliferation activity and poor immune microenvironment in Cluster3 subgroup were associated with its poor prognosis, larger tumor size, high AFP levels, high incidence of tumor thrombus and higher clinical stage. The characteristics of the Cluster2 subgroup were between the Cluster1 and Cluster3 groups. In addition, MCM2-7, RFC2-5, MSH2, MSH6, SMC2, SMC4, NCPAG and TOP2A proteins were significantly upregulated in the Cluster3 subgroup. Meanwhile, abnormally high phosphorylation levels of these proteins were associated with high levels of DNA repair, telomere maintenance and proliferative features. Therefore, these proteins could be identified as potential diagnostic and prognostic markers. In general, our research provided a novel analytical protocol and insights for the robust classification, treatment and prevention of HBV-infected HCC.

Proliferative characteristics of eutopic and ectopic endometrium in adenomyosis using AgNOR technology

Adenomyosis is a benign lesion of the uterus, which is characterized by the presence and pathological growth of endometrial glands and stroma in the myometrium. During adenomyosis, endometrial tissue abnormally grown in the myometrium is called ectopic endometrium, and endometrial tissue in the uterine cavity is called eutopic endometrium. Women with adenomyosis may experience abnormal uterine bleeding, dysmenorrhea, dyspareunia, and infertility, although, in one-third of women, the lesion is asymptomatic. Adenomyosis is the most frequently diagnosed pathology after hysterectomy in perimenopausal women due to heavy bleeding or pain. AgNOR is used as an aid for evaluating the proliferative activity of cells. Certain types of studies have demonstrated their effectiveness in differentiating various benign or malignant processes, as well as in determining the histological grade of intraepithelial neoplastic processes. Within the framework of this study, the proliferative activity of the eutopic and ectopic endometrium, as well as their stroma and normal myometrium, was evaluated on archival blocks fixed in formalin and embedded in paraffin (FFPE), diagnosed in cases of adenomyosis, using AgNOR technology. In the studied system, eutopic endometrium - basal endometrium - in both the glandular and stromal components of ectopic endometrium, the highest proliferative activity is manifested in ectopic endometrium, which is why it can probably represent a risk for the development of neoplasms.

IPSC-Derived MSCs Are a Distinct Entity of MSCs with Higher Therapeutic Potential than Their Donor-Matched Parental MSCs

Mesenchymal stromal cells derived from induced pluripotent stem cells (iMSCs) have been proposed as alternative sources of primary MSCs with various advantages for cell therapeutic trials. However, precise evaluation of the differences between iMSCs and primary MSCs is lacking due to individual variations in the donor cells, which obscure direct comparisons between the two. In this study, we generated donor-matched iMSCs from individual bone marrow-derived MSCs and directly compared their cell-autonomous and paracrine therapeutic effects. We found that the transition from primary MSCs to iMSCs is accompanied by a functional shift towards higher proliferative activity, with variations in differentiation potential in a donor cell-dependent manner. The transition from MSCs to iMSCs was associated with common changes in transcriptomic and proteomic profiles beyond the variations of their individual donors, revealing expression patterns unique for the iMSCs. These iMSC-specific patterns were characterized by a shift in cell fate towards a pericyte-like state and enhanced secretion of paracrine cytokine/growth factors. Accordingly, iMSCs exhibited higher support for the self-renewing expansion of primitive hematopoietic progenitors and more potent immune suppression of allogenic immune responses than MSCs. Our study suggests that iMSCs represent a separate entity of MSCs with unique therapeutic potential distinct from their parental MSCs, but points to the need for iMSC characterization in the individual basis.

Novel neuron-Schwann cell co-culture models to study peripheral nerve degeneration and regeneration.

Novel neuron-Schwann cell co-culture models to study peripheral nerve degeneration and regeneration.

Role of the NF-kB/parkin/vegfr-1 pathway associated with hypoxic-ischemic insult in germinal matrix samples of newborn infants.

Given the high proliferative activity of germinal matrix and its direct correlation with hypoxemia, it is necessary to investigate the possible molecular regulation pathways, to understand the existing clinical relationship between the hypoxic-ischemic insult and the biomarkers NF-kB, AKT-3, Parkin, TRK-C and VEGFR-1. A hundred and eighteen germinal matrix samples of the central nervous system of patients who died in the first 28 days of life were submitted to histological and immunohistochemistry analysis to identify the tissue immunoexpression of those biomarkers related to asphyxia, prematurity, and death events within 24h. A significantly increased tissue immunoexpression of NF-kB, AKT-3 and Parkin was observed in the germinal matrix of preterm infants. In addition, significantly decreased tissue immunoexpression of VEGFR-1 and NF-kB was observed in patients who experienced asphyxia followed by death within 24 hours. The results suggest a direct involvement between the hypoxic-ischemic insult and NF-kB and VEGFR-1 markers since a decreased immunoexpression of these biomarkers was observed in asphyxiated patients. Furthermore, it is suggested that there was not enough time for VEGFR-1 to be transcribed, translated and expressed on the surface of the plasma membrane. This temporality can be observed in the relationship between NF-kB expression and the survival time of individuals who died within 24 hours, suggesting that this factor is essential for the production of VEGFR-1 and, therefore, to carry out the necessary remodeling effect to neovascularize the affected region.

Long term trends of breast cancer incidence according to proliferation status

BackgroundLong-term breast cancer incidence trends according to proliferation status are poorly described. We studied time-trends in breast cancer incidence, using mitotic count and Ki-67 as markers of proliferation.MethodsAmong 83,298 Norwegian women followed for breast cancer occurrence 1961–2012, 2995 incident breast cancers were diagnosed. Ki-67 was assessed using immunohistochemistry on tissue microarrays and mitoses were counted on whole sections. We compared incidence rates according to proliferation status among women born 1886–1928 and 1929–1977, estimating age-specific incidence rate ratios. We performed multiple imputations to account for unknown proliferation status. Mean values of Ki-67 and mitotic counts were calculated, according to age and birth year. We performed separate incidence analyses for HER2+ and triple negative breast cancers.ResultsAmong women aged 40–69 years, incidence rates of tumours with low-proliferative activity were higher among those born in 1929 or later, compared to before 1929, according to Ki-67 and mitotic count. Incidence rates of tumours with high-proliferative activity were also higher in women born in 1929 or later compared to before 1929 according to Ki-67, but not according to mitotic count. Mean values of Ki-67 and mitotic count varied according to age and birth year. In subtype-specific analyses we found an increase of high-proliferative HER2+ tumours according to Ki-67 in women born in 1929 or later, compared to before 1929.ConclusionsThere has been a temporal increase in both low- and high-proliferative breast cancers.

A UNIQUE ADIPOCYTE PROGENITOR POPULATION PROMOTES AGE-RELATED ADIPOSITY

Abstract The average fat mass in adults increases dramatically with age, and older people often suffer from visceral obesity and related adverse metabolic disorders. Unfortunately, how aging leads to fat accumulation is poorly understood. It is known that fat cell (adipocyte) turnover is very low in young mice, similar to that in young humans. Here, we find that mice mimic age-related fat expansion in humans. In vivo lineage tracing shows that massive adipogenesis (the generation of new adipocytes), especially in the visceral fat, is triggered during aging. Thus, in contrast to most types of adult stem cells that exhibit a reduced ability to proliferate and differentiate, the adipogenic potential of adipocyte progenitor cells (APCs) is unlocked by aging. In vivo transplantation and 3D imaging of transplants show that APCs in aged mice cell-autonomously gain high adipogenic capacity. Single-cell RNA sequencing analyses reveal that aging globally remodels APCs. Herein, we identify a novel committed preadipocyte population that is age-specific (CP-A), existing both in mice and humans, with a global activation of proliferation and adipogenesis pathways. CP-A cells display high proliferation and adipogenesis activity, both in vivo and in vitro. LIFR serves as a functional maker of CP-A, which promotes CP-A proliferation. Together, these findings define a new fundamental mechanism involved in fat tissue aging and offer prospects for preventing and treating age-related metabolic disorders.

Функціональна активність гемопоетичних клітин-попередників кордової крові у довготривалій культурі на гідрогелевій підложці in vitro

Development of new and improvement of existing models of long-term cultivation of hematopoietic stem cells (HSC) and cord blood progenitor cells is one of the most important directions in modern cell biology, related to obtaining a sufficient amount of hematopoietic tissue for experimental and clinical use. The aim of the work was to investigate the functional activity of hematopoietic progenitor cells from cord blood during long-term cultivation on a soft polyacrylamide gel substrate in the presence of a cytokine complex and to determine the advantages of this method of maintaining hematopoiesis for further use in transplantation. To realize this purpose, the following methods were used: the method of long-term cultivation in vitro on a hydrogel substrate, the method of colony formation in semi-liquid agar, cytological research methods, light and inverted microscopy, statistical research methods. Due to the use of the hydrogel substrate together with the cytokine complex, a high proliferative activity of hematopoietic cells is observed, which is reflected in their expansion during long-term cultivation (5 weeks), as well as of high colony-forming activity (521.5 ± 7.5.105 per explanted cells). Thus, the stiffness of the substrate must be taken into account for the expansion of stem cells and their immediate descendants. The presence of a soft substrate made of polyacrylamide gel along with a complex of cytokines ensures the expansion of hematopoietic cells due to the long- term support of hematopoiesis.

Highly recurrent BRAF p.V595E mutation in canine papillary oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is the most common oral epithelial malignancy in dogs. It exhibits locally aggressive biological behaviour with the potential to metastasize, and a reported 1-year survival rate of 0% when left untreated. Expression studies suggest that aberrant MAPK signalling plays a key role in canine OSCC tumorigenesis, which is consistent with BRAF and HRAS MAPK-activating mutations reported in some tumours. Several morphological subtypes of canine OSCC have been described, with papillary, conventional, and basaloid as the most common patterns. We hypothesized that mutational differences may underlie these phenotypic variations. In this study, targeted Sanger sequencing and restriction fragment length polymorphism assays demonstrate that up to 85.7% of canine papillary OSCC (n=14) harbour a BRAF p.V595E mutation. Assessment of neoplastic epithelial cell proliferation using Ki67 immunolabelling (n=10) confirmed a relatively high proliferation activity, consistent with their known aggressive clinical behaviour. These findings underscore a consistent genetic feature of canine papillary OSCC and provide a basis for the development of novel diagnostic and targeted therapeutic approaches that can improve the quality of veterinary care.

Tumor biology: molecular genetics mechanisms of initiation, promotion and progression

For all tumors, regardless of type and tissue, there are a number of common features. These include high proliferative activity, disturbances in the mechanisms of apoptosis and "invisibility" for the cells of the immune system. At the basis of all these disorders lies a malfunction in the interaction between signaling proteins inside the cell, due to a violation of the expression of genes responsible for their synthesis. The aim of the review is to study the molecular biological mechanisms of carcinogenesis, characteristics and features of tumor cells that affect the development of the disease, progression, drug resistance and clinical results and the possibilities of targeted therapy of oncological diseases. Material and methods. The search for literature sources was carried out using PubMed, CyberLeninka, eLibrary, Google Scholar databases; the search depth was 15 years. Results. Recent works have shown the evolution of views on the classification of tumors from the point of view of the mechanisms of damage to the processes of cellular regulation and histogenesis, as well as the development of the modern theory of carcinogenesis, ideas about the mechanisms of development of tumor diseases and the prospects of modern therapies. Conclusion. The study of molecular biological mechanisms of carcinogenesis, characteristics and features of tumor cells is not only fundamental, but also of applied importance in order to predict the development of the disease, progression, drug resistance, clinical results and the possibilities of targeted therapy.

Immunohistochemical study of neural stem cell lineage markers in canine brains, gliomas, and a glioma cell line.

Neural stem cells (NSCs) produce neuron intermediate progenitor cells (nIPC), oligodendrocyte precursor cells (OPCs), and immature astrocytes. To confirm NSC lineages in the normal canine brain and the association of these cells with gliomas, an immunohistochemical study was conducted on fetal and adult canine brains, gliomas, and a glioma cell line. In fetal brains, glial fibrillary acidic protein (GFAP)- and nestin-immunolabeled NSC were observed in the ventricular zone, β-3 tubulin- and/or neuronal nuclei (NeuN)-immunolabeled nIPC in the subventricular zone (SVZ), and platelet-derived growth factor receptor-α (PDGFR-α)- and OLIG2-immunolabeled OPC and GFAP- and OLIG2-immunolabeled immature astrocytes in the SVZ and intermediate zone. Ki-67 immunohistochemistry revealed that nIPC exhibited high proliferative activity. Quiescent nIPC and OPC were observed in adult brains. Among 58 glioma cases including 4 low-grade oligodendrogliomas (LGOGs), 48 high-grade oligodendrogliomas (HGOGs), 1 low-grade astrocytoma, and 5 high-grade astrocytomas (HGACs), immunohistochemical analyses revealed that oligodendrogliomas expressed PDGFR-α and OLIG2, whereas astrocytomas expressed GFAP and OLIG2. HGOG showed significantly higher immunohistochemical scores for NeuN and β-3 tubulin than LGOG. The Ki-67 labeling index was high in PDGFR-α and NeuN-immunolabeled tumor cells, and low in β-3 tubulin- and synaptophysin-immunolabeled cells. A HGOG cell line possessed the same immunohistochemical characteristics as HGOG. In this study, glioma cells with the OPC and IPC immunophenotypes had a higher Ki-67 labeling index, indicating their high proliferative activity. Furthermore, high-grade gliomas showed the characteristics of nIPC and neurons, which may suggest the pluripotent NSC lineage nature of these tumors.

R-DA-Edoch/R-DHAP Alteration Could Overcome High Risk Factors in Younger Mantle Cell Lymphoma

Background: Mantle-cell lymphoma (MCL) is a rare subtype of non-Hodgkin's lymphoma, with a median overall survival of only 3~5 years in the era of immunochemotherapy. Although first-line intensive chemotherapy with rituximab (R) and high dose cytarabine combined with autologous transplantation (ASCT) has significantly prolonged the survival of patients with MCL, some patients still die within few years. Our published data show that Chinese patients with MCL are younger and have a higher proportion of TP53 deletion and MYC abnormality [Oncotarget, 2015 Dec 8;6(39):42362-71]. Considering the efficacy and tolerability of the dose-enhanced immunochemotherapy R-DA-EDOCH regimen in patients with multiple high-risk aggressive B-cell lymphomas and the potential higher proliferative activity of tumor cells in Chinese patients, we conducted a prospective clinical study of alternate R-DA-EDOCH/R-DHAP induction therapy for young patients with newly-diagnosed MCL (NCT02858804). Here, we analyzed the efficacy and safety of this regimen. Methods: The main inclusion criteria for patients with newly-diagnosed MCL included age ≤ 65 years, Ann Arbor stage II~IV and ECOG ≤ 1. During induction, patients received alternate R-DA-EDOCH/R-DHAP for two cycles. Patients who achieved less than partial remission would quit the trial, while the other patients would continue another one cycle of treatment. After finishing the 3 cycles of inductive treatment, patients would decide whether to continue ASCT consolidation therapy or another cycle of consolidation. R maintenance therapy was recommended to all patients. The primary endpoint of this study was the progression-free survival (PFS). Key secondary endpoints were complete response rate (CRR) and objective response rate (ORR) at the end of induction (EOI), overall survival (OS), minimal residual disease (MRD) status and the incidence of adverse events (AEs). Results: In this study, we recruited 55 patients with a median age of 53 years (range, 37~65 years) and a male/female ratio of 2.2:1. Totally, 61.8% of patients were of at least one risk factor (HR) according to risk stratification, pathological classification and molecular genetic abnormalities. All patients received at least two cycles of alternate R-DA-EDOCH/R-DHAP immunochemotherapy, and one patient dropped out due to the poor response. The ORR of the alternate R-DA-EDOCH/R-DHAP inductive treatment was 98% (95% CI 90-100), and the CRR was 89% (95%CI 78-96). There was no significant difference in CRR and ORR among patients with different prognostic characteristics. Moreover, patients rapidly achieved MRD-negative and the rate of MRD-negative after 2 cycles of treatment, the end of the inductive therapy and the end of the overall treatment was 79.2%, 88.7% and 90.6%, respectively. The median follow-up of the cohort was 31 months (range 4-86). The 3-year PFS and OS rates were 66.3% and 85.4%, respectively. Sequential ASCT consolidation and/or R maintenance slightly increased CRR but significantly prolonged PFS of patients (3-year PFS rate: ASCT+R 90.4% versus R 64.6% versus without ASCT/R 33.3%, P<0.001). There was no significance difference in PFS and OS between patients with (n=34) and without (n=9) HR (PFS P=0.537, OS P=0.408). For patients receiving ASCT and R maintenance, OS was even longer in patients with HR (mOS not reach versus 47m, P=0.007). Univariate analysis showed that age≥60 years, blastoid/pleomorphic pathologic morphology and TP53 double-allele events were still the risk factors for PFS in this therapeutic pattern. However, high tumor proliferative activity (such as Ki-67≥30% and MYC abnormality) and specific genetic abnormalities (such as 17p deletion, 9p deletion, etc) showed no significant adverse prognostic influence. In particular, patients who achieved MRD-negative by treatment had longer PFS (P<0.001). The alternate R-DA-EDOCH/R-DHAP regimen was well tolerated, and no patients were discontinued due to intolerance of treatment-related AEs. AEs were mainly hematologic and at least one hematologic AEs occurred in 85.5% of the patients. Grade 3-4 leukopenia and neutropenia were found in 49.1% and 58.2% of the patients, respectively. Conclusion: The alternate R-DA-EDOCH/R-DHAP induction therapy showed great efficacy and tolerability in young patients with newly diagnosed MCL and could overcome the adverse prognostic impact of some high-risk factors. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Adhesive Bioinspired Coating for Enhancing Glass-Ceramics Scaffolds Bioactivity.

Bioceramic scaffolds, composed of a biphasic composite containing bioactive glass and hydroxyapatite, were prepared in this work to overcome the intrinsic limits of the two components taken separately (in particular, their specific reactivities and dissolution rates, which should be tunable as a function of the given clinical requirements). To mimic the biological environment and tune the different stages of cellular response, a coating with gelatin and chondroitin sulphate via Layer-by-Layer (LbL) assembly was presented and discussed. The resulting functionalized scaffolds were affected by the coating in terms of microstructure and porosity. In addition, the LbL coating significantly enhanced the seeded cell behaviour, with high adhesion, proliferation and osteogenic activity, as revealed by the alkaline phosphatase activity and overexpression of osteopontin and osteocalcin.

The number of the intraepithelial T cells correlate with the proliferation index in human bulbourethral gland epithelium

BackgroundOur study has immunohistochemically examined T cells localization and number as well as proliferative activity level for the bulbourethral gland epithelium in men of different ages, using monoclonal antibodies against CD45RO and proliferating cell nuclear antigen (PCNA). ResultsWe have found that the T cells have been localized mainly in excretory ducts epithelium of the glands in any age group, meanwhile their relative number varies with age. The excretory ducts epithelium has shown a high proliferative activity when in acini the PCNA index has been low. Postnatal dynamics of the epithelium proliferative activity positively correlates with age-related density fluctuations in lymphocytic infiltration of the glands. ConclusionsWe consider that intraepithelial T cells may contribute to the regulation of epithelial cells proliferation in the bulbourethral glands.

DNMT3A R882H mutation drives daunorubicin resistance in acute myeloid leukemia via regulating NRF2/NQO1 pathway

BackgroundDNA methyltransferase 3A (DNMT3A) often mutate on arginine 882 (DNMT3AR882) in acute myeloid leukemia (AML). AML patients with DNMT3A R882 mutation are usually resistant to daunorubicin treatment; however, the associated mechanism is still unclear. Therefore, it is urgent to investigate daunorubicin resistance in AML patients with DNMT3A R882 mutant.MethodAML cell lines with DNMT3A-wild type (DNMT3A-WT), and DNMT3A-Arg882His (DNMT3A-R882H) mutation were constructed to investigate the role of DNMT3A R882H mutation on cell proliferation, apoptosis and cells’ sensitivity to Danunorubin. Bioinformatics was used to analyze the role of nuclear factor-E2-related factor (NRF2) in AML patients with DNMT3A R882 mutation. The regulatory mechanism of DNMT3A R882H mutation on NRF2 was studied by Bisulfite Sequencing and CO-IP. NRF2 inhibitor Brusatol (Bru) was used to explore the role of NRF2 in AML cells carried DNMT3A R882H mutation.ResultsAML cells with a DNMT3A R882H mutation showed high proliferative and anti-apoptotic activities. In addition, mutant cells were less sensitive to daunorubicin and had a higher NRF2 expression compared with those in WT cells. Furthermore, the NRF2/NQO1 pathway was activated in mutant cells in response to daunorubicin treatment. DNMT3A R882H mutation regulated the expression of NRF2 via influencing protein stability rather than decreasing methylation of NRF2 promoter. Also, NRF2/NQO1 pathway inhibition improved mutant cells’ sensitivity to daunorubicin significantly.ConclusionOur findings identified NRF2 as an important player in the regulation of cell apoptosis through which helps mediate chemoresistance to daunorubicin in AML cells with DNMT3A R882H mutation. Targeting NRF2 might be a novel therapeutic approach to treat AML patients with a DNMT3A R882H mutation.9jD1n7ooVY8uHe1s9Su8KLVideo abstract