Inhibition of glioblastoma and enhancement of survival via the use of mibefradil in conjunction with radiosurgery

Object The survival of patients with high-grade gliomas remains unfavorable. Mibefradil, a T-type calcium channel inhibitor capable of synchronizing dividing cells at the G1 phase, has demonstrated potential benefit in conjunction with chemotherapeutic agents for gliomas in in vitro studies. In vivo study of mibefradil and radiosurgery is lacking. The authors used an intracranial C6 glioma model in rats to study tumor response to mibefradil and radiosurgery. Methods Two weeks after implantation of C6 cells into the animals, each rat underwent MRI every 2 weeks thereafter for 8 weeks. After tumor was confirmed on MRI, the rats were randomly assigned to one of the experimental groups. Tumor volumes were measured on MR images. Experimental Group 1 received 30 mg/kg of mibefradil intraperitoneally 3 times a day for 1 week starting on postoperative day (POD) 15; Group 2 received 8 Gy of cranial radiation via radiosurgery delivered on POD 15; Group 3 underwent radiosurgery on POD 15, followed by 1 week of mibefradil; and Group 4 received mibefradil on POD 15 for 1 week, followed by radiosurgery sometime from POD 15 to POD 22. Twenty-seven glioma-bearing rats were analyzed. Survival was compared between groups using Kaplan-Meier methodology. Results Median survival in Groups 1, 2, 3, and 4 was 35, 31, 43, and 52 days, respectively (p = 0.036, log-rank test). Two animals in Group 4 survived to POD 60, which is twice the expected survival of untreated animals in this model. Analysis of variance and a post hoc test indicated no tumor volume differences on PODs 15 and 29. However, significant volume differences were found on POD 43; mean tumor volumes for Groups 1, 2, 3, and 4 were 250, 266, 167, and 34 mm3, respectively (p = 0.046, ANOVA). A Cox proportional hazards regression test showed survival was associated with tumor volume on POD 29 (p = 0.001) rather than on POD 15 (p = 0.162). In vitro assays demonstrated an appreciable and dose-dependent increase in apoptosis between 2- and 7-μM concentrations of mibefradil. Conclusions Mibefradil response is schedule dependent and enhances survival and reduces glioblastoma when combined with ionizing radiation.

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Combination of 177lutetium-Satetraxetan-Lilotomab and Rituximab Results in Improved Therapeutic Effect in Preclinical Models of Non-Hodgkin Lymphoma

Blood ◽

10.1182/blood.v128.22.4189.4189 ◽

2016 ◽

Vol 128 (22) ◽

pp. 4189-4189

Author(s):

Ada H.V. Repetto-Llamazares ◽

Roy Hartvig Larsen ◽

Adam O'Shea ◽

Jostein Dahle

Keyword(s):

Therapeutic Effect ◽

Combination Treatment ◽

Tumor Volume ◽

Proportional Hazards ◽

Cox Proportional Hazards ◽

Employment Equity ◽

Non Hodgkin Lymphoma ◽

Equity Ownership

Abstract Aim: To investigate the therapeutic effect of combined treatment with the anti-CD37 antibody radionuclide conjugate (ARC) 177Lu-satetraxetan-lilotomab (177Lu-p-SCN-Bn-DOTA-HH1, tradename Betalutin®, short name 177Lu-lilotomab) and the anti-CD20 immunotherapy with rituximab in a subcutaneous (s.c.) preclinical model of non-Hodgkin lymphoma (NHL). These studies build on previously presented data that showed that treatment with 177Lu-lilotomab increased binding of rituximab in NHL cells in-vitro and in-vivo. Materials and Methods:The therapeutic effect of the combination of 177Lu-lilotomab and rituximab was studied in nude mice with s.c. Burkitt's lymphoma (Daudi) xenografts using 250 MBq/kg 177Lu-lilotomab, 1 dose of 40 mg/kg rituximab, 4 doses of 10 mg/kg rituximab orNaCl. Treatmentswere given every 3 to 4 days (Table 1). Nine to 10 mice bearinga total of 16 to 18 tumors per group were used in the study. Tumor volume was measured every 2-3 days and weekly after study day 100. The therapeutic effect of the combination treatmentswas compared with the therapeutic effect of each of the treatments alone using Log-Rank and clustered cox-proportional hazards tests. Results:The effect of the combination treatment in mice with s.c. xenografts was better than that of each treatment alone (Figure 1 & 2). The median survival of mice given the combination treatment was longer (>157 days) than each of the treatments alone (31-60 days) and of the sum of both treatments alone (91-100 days), but the difference was not statistically significant (Table 2). The study is currently on-going and none of the mice given the combination treatment have any palpable tumors, so it is expected that the median survival of those mice eventually will double the sum of both treatments alone. Median time to quadruplicate tumor volume was 2 times longer for the combination treatments (>157 days) than in any of the treatments alone (25-49 days) and longer than the sum of both treatments alone (74-82 days) (Table 2). When using time to quadruplicate tumor volume as end-point, the 177Lu-lilotomab + 4 x rituximab combination was significantly different from either treatment alone (p < 0.01, clustered cox-proportional hazards model). Moreover, the median time to complete remission of tumors in mice given the combination treatment was 5 times shorter (13 days) than each of the treatments alone (>66 days) (p<0.05). Three mice in the combination treatment of 177Lu-lilotomab + 4 x rituximab and one mouse in the combination treatment of 177Lu-lilotomab + 1 x rituximab doses were sacrificed due to body weight loss 14, 110, 118 and 157 days after injection of 177Lu-lilotomab respectively. It is not known if this was normal age-related symptoms or random occurrences or due to treatment toxicity. Histopathology examination will be included in the next mice tobe sacrificed to study this in more detail and evaluate if there is any late toxicity due to the combination treatment. Conclusion:Treatment of NHL in-vitro and in-vivo with 177Lu-lilotomab results in an increased binding, tumor uptake and tumor suppression of anti-CD20 immunotherapy. If the same effect is confirmed in clinical studies, it could change the way ARC and CD20 immunotherapy would be used in the future. Disclosures Repetto-Llamazares: Nordic Nanovector ASA: Employment, Equity Ownership. Larsen:Nordic Nanovector ASA: Equity Ownership. O'Shea:Nordic Nanovector ASA: Employment, Equity Ownership. Dahle:Nordic Nanovector ASA: Employment, Equity Ownership.

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Mutational Analysis of the Chemoreceptor-Coupling Domain of the Escherichia coli Chemotaxis Signaling Kinase CheA

Journal of Bacteriology ◽

10.1128/jb.188.9.3299-3307.2006 ◽

2006 ◽

Vol 188 (9) ◽

pp. 3299-3307 ◽

Cited By ~ 25

Author(s):

Jinshi Zhao ◽

John S. Parkinson

Keyword(s):

Escherichia Coli ◽

Mutational Analysis ◽

In Vitro Assays ◽

Mutant Proteins ◽

Receptor Coupling ◽

Group 4 ◽

Group 2 ◽

Group 1

ABSTRACT During chemotactic signaling by Escherichia coli, autophosphorylation of the histidine kinase CheA is coupled to chemoreceptor control by the CheW protein, which interacts with the C-terminal P5 domain of CheA. To identify P5 determinants important for CheW binding and receptor coupling control, we isolated and characterized a series of P5 missense mutants. The mutants fell into four phenotypic groups on the basis of in vivo behavioral and protein stability tests and in vitro assays with purified mutant proteins. Group 1 mutants exhibited autophosphorylation and receptor-coupling defects, and their CheA proteins were subject to relatively rapid degradation in vivo. Group 1 mutations were located at hydrophobic residues in P5 subdomain 2 and most likely caused folding defects. Group 2 mutants made stable CheA proteins with normal autophosphorylation ability but with defects in CheW binding and in receptor-mediated activation of CheA autophosphorylation. Their mutations affected residues in P5 subdomain 1 near the interface with the CheA dimerization (P3) and ATP-binding (P4) domains. Mutant proteins of group 3 were normal in all tests yet could not support chemotaxis, suggesting that P5 has one or more important but still unknown signaling functions. Group 4 mutant proteins were specifically defective in receptor-mediated deactivation control. The group 4 mutations were located in P5 subdomain 1 at the P3/P3′ interface. We conclude that P5 subdomain 1 is important for CheW binding and for receptor coupling control and that these processes may require substantial motions of the P5 domain relative to the neighboring P3 and P4 domains of CheA.

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Association of gender, age, and ethnicity with survival in patients with pancreas cancer

Journal of Clinical Oncology ◽

10.1200/jco.2009.27.15_suppl.e15587 ◽

2009 ◽

Vol 27 (15_suppl) ◽

pp. e15587-e15587

Author(s):

M. Ottochian ◽

D. Yang ◽

A. El-Khoueiry ◽

S. Iqbal ◽

A. Pohl ◽

...

Keyword(s):

Pancreatic Cancer ◽

Proportional Hazards ◽

Patient Characteristics ◽

The United States ◽

Lymph Node Involvement ◽

Cox Proportional Hazards ◽

Age At Menarche ◽

Age Group

e15587 Background: Pancreatic cancer (PC) is the fourth leading cause of cancer death in the United States. However little is still known about factors that influence its development and progression. Recent data suggest that PC is, at least in part, an estrogen- dependent disease; there is growing epidemiological evidence that aspects of reproductive history and hormonal exposure are associated with risk of this disease. It was shown that age at menarche of <13 is associated with less risk of PC. However no data are available whether gender is associated with outcome in patients with PC. The purpose of this study was to test whether age, gender or ethnicity influence the outcome in PC. Methods: The data of the 50,302 adults diagnosed with PC between 1988 and 2004 were extracted from the Surveillance Epidemiology and End Results public use database. These included 24,240 patients diagnosed with localized pancreatic cancer (LPC) and 26,062 patients with metastatic pancreatic cancer (MPC). Demographic, clinical variables and survival time were retrieved. The primary endpoint was overall survival. We constructed Cox proportional hazards models to evaluate association between patient characteristics and survival in LPC and MPC separately. Pair interactions were also tested. Results: On multivariate analysis gender, age, race, marital status, tumor size, grade, histology, type of treatment and lymph node involvement were found to be independent predictors of survival. Females had a significant longer survival, with an HR of 0.959 (95% CI: 0.932–0.987) among patients with LPC and an HR of 0.918 (95%CI: 0.894–0.942) among patients with MPC. Each age group displayed a significant longer survival than its correspondent older age group. When we combined age and gender in the analysis, females had a longer survival than males in each single age group in the MPC group. In the LPC group the longer survival of female patients was only observed in the youngest age group. Conclusions: This is the first and largest study to address gender and outcome in PC. Our data suggest that the estrogen pathway may play an important prognostic role in patient with this disease. These data also warrant further in vitro and in vivo investigations on the mechanisms of estrogen and pancreas progression. No significant financial relationships to disclose.

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Connecting METTL3 and intratumoural CD33+ MDSCs in predicting clinical outcome in cervical cancer

Journal of Translational Medicine ◽

10.1186/s12967-020-02553-z ◽

2020 ◽

Vol 18 (1) ◽

Author(s):

Huan-he Ni ◽

Lin Zhang ◽

He Huang ◽

Shu-qin Dai ◽

Jiang Li

Keyword(s):

Cervical Cancer ◽

Proportional Hazards ◽

Cox Model ◽

Disease Free Survival ◽

Suppressor Cells ◽

Cox Proportional Hazards ◽

Tumour Cells ◽

Rank Test ◽

Independent Factor

Abstract Background Methyltransferase-like 3 (METTL3) is a member of the m6A methyltransferase family and acts as an oncogene in cancers. Recent studies suggest that host innate immunity is regulated by the enzymes controlling m6A epitranscriptomic changes. Here, we aim to explore the associations between the levels of METTL3 and CD33+ myeloid-derived suppressor cells (MDSCs) in tumour tissues and the survival of patients with cervical cancer (CC). Methods Specimens of paraffin embedded tumour from 197 CC patients were collected. The expression levels of METTL3 and CD33 were measured by immunohistochemical (IHC) staining. The clinical associations of the IHC variants were analysed by Pearson’s or Spearman’s chi-square tests. Overall survival (OS) and disease-free survival (DFS) were estimated by the Kaplan–Meier method and log-rank test. Hazard ratios (HRs) and independent significance were obtained via Cox proportional hazards models for multivariate analyses. METTL3 in CD33+ cells or CC-derived cells was knocked down by METTL3-specific siRNA, and MDSC induction in vitro was performed in a co-culture system in the presence of METTL3-siRNA and METTL3-knockdown-CC-derived cells compared with that of the corresponding controls. Results We found that tumour tissues displayed increased levels of METTL3 and CD33+ MDSCs compared with tumour-adjacent tissues from the same CC patients. Importantly, METTL3 expression was positively related to the density of CD33+ cells in tumour tissues (P = 0.011). We further found that the direct CD33+CD11b+HLA-DR− MDSC induction and tumour-derived MDSC induction in vitro were decreased in the absence of METTL3. The level of METTL3 in tumour microenvironments was significantly related to advanced tumour stage. The levels of METTL3 and CD33+ MDSCs in tumour tissues were notably associated with reduced DFS or OS. Cox model analysis revealed that the level of METTL3 in tumour cells was an independent factor for patient survival, specifically for DFS (HR = 3.157, P = 0.022) and OS (HR = 3.271, P = 0.012), while the CD33+ MDSC number was an independent predictor for DFS (HR: 3.958, P = 0.031). Interestingly, in patients with advanced-disease stages (II–IV), METTL3 in tumour cells was an independent factor for DFS (HR = 6.725, P = 0.010) and OS (HR = 5.140, P = 0.021), while CD33+ MDSC density was an independent factor for OS (HR = 8.802, P = 0.037). Conclusion Our findings suggest that CD33+ MDSC expansion is linked to high levels of METTL3 and that METTL3 and CD33+ MDSCs are independent prognostic factors in CC.

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Reduced LH sensitivity in vivo and in vitro of corpora lutea induced during anoestrus by GnRH, and during the late breeding season, in Scottish Blackface ewes

Journal of Endocrinology ◽

10.1677/joe.1.05918 ◽

2004 ◽

Vol 183 (3) ◽

pp. 517-526 ◽

Cited By ~ 2

Author(s):

T A Bramley ◽

D Stirling ◽

G S Menzies ◽

D T Baird

Keyword(s):

Breeding Season ◽

High Plasma ◽

Corpora Lutea ◽

Plasma Progesterone ◽

Group 4 ◽

Group 3 ◽

Group 2 ◽

Group 1

Scottish Blackface ewes were synchronised in mid-breeding (November; group 1; n=12 ewes) or late-breeding season (March; group 2; n=16). Anoestrous ewes (May) were treated with progestagen sponges for 7 days and then given 250 ng GnRH 3-hourly for 24 h, 2-hourly for 24 h and hourly for a further 24 h (group 3; n=12). A second group of anoestrous ewes (group 4, n=19) received three bolus injections (30 μg) of GnRH at 90-min intervals without progestagen pretreatment. After ovulation, ewes were bled twice daily until slaughter (day 4 or day 12: oestrus=day 0). Mid-breeding season (group 1) and anoestrous ewes in group 3 formed ‘adequate’ corpora lutea (CL) with high plasma progesterone levels (3–4 ng/ml) maintained for at least 12 days, and responded in vivo to ovine LH (oLH) (10 μg) with a rise in plasma progesterone on day 11 (group 3, but not group 1, ewes also responded on day 3). CL minces from these ewes responded to human chorionic gonadotrophin (hCG) in vitro with a dose-dependent increase in progesterone secretion. Ewes in group 4 had a foreshortened luteal phase (8–10 days) and low plasma progesterone levels (~1 ng/ml), consistent with formation of inadequate CL. LH injection failed to induce a significant plasma progesterone increase. Furthermore, although progesterone secretion in vitro in response to maximally stimulating doses of hCG or dibutyryl cAMP (dbcAMP) was similar to that in adequate CL, the sensitivity of these CL to hCG (EC (effective concentration)50, 1 IU hCG/ml) was reduced 10-fold compared with adequate CL (EC50, 0.1 IU hCG/ml; P<0.01). Ewes that ovulated in the late breeding season (group 2) had high plasma progesterone, although levels began to decrease after day 10. Injection of oLH in vivo increased plasma progesterone. However, sensitivity to hCG in vitro (EC50, 0.5 IU hCG/ml) was intermediate between that of adequate luteal tissue (groups 1 and 3; EC50, 0.1 IU/ml) and that of group 4 ewes (EC50, 1 IU hCG/ml). Our data demonstrate a markedly reduced luteal sensitivity to LH in vivo and hCG in vitro in Scottish Blackface ewes with inadequate CL, and suggest that a similar loss of sensitivity to LH may occur in the late breeding season.

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Garlic Consumption and All-Cause Mortality among Chinese Oldest-Old Individuals: A Population-Based Cohort Study

Nutrients ◽

10.3390/nu11071504 ◽

2019 ◽

Vol 11 (7) ◽

pp. 1504 ◽

Cited By ~ 1

Author(s):

Shi ◽

Lv ◽

Mao ◽

Yuan ◽

Yin ◽

...

Keyword(s):

Proportional Hazards ◽

Oldest Old ◽

Experimental Studies ◽

Population Based ◽

Cox Proportional Hazards ◽

Sensitivity Analyses ◽

Protective Effects ◽

All Cause Mortality

In vitro and in vivo experimental studies have shown garlic has protective effects on the aging process; however, there is no evidence that garlic consumption is associated with all-cause mortality among oldest-old individuals (≥80 years). From 1998 to 2011, 27,437 oldest-old participants (mean age: 92.9 years) were recruited from 23 provinces in China. The frequencies of garlic consumption at baseline and at age 60 were collected. Cox proportional hazards models adjusted for potential covariates were constructed to estimate hazard ratios (HRs) relating garlic consumption to all-cause mortality. Among 92,505 person-years of follow-up from baseline to September 1, 2014, 22,321 participants died. Participants who often (≥5 times/week) or occasionally (1–4 times/week) consumed garlic survived longer than those who rarely (less than once/week) consumed it (p < 0.001). Participants who consumed garlic occasionally or often had a lower risk for mortality than those who rarely consumed garlic at baseline; the adjusted HRs for mortality were 0.92(0.89–0.94) and 0.89(0.85–0.92), respectively. The inverse associations between garlic consumption and all-cause mortality were robust in sensitivity analyses and subgroup analyses. In this study, habitual consumption of garlic was associated with a lower all-cause mortality risk; this advocates further investigation into garlic consumption for promoting longevity.

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71 WILL DEHYDRATION BY EXPOSURE TO SUCROSE IMPROVE POST-VITRIFICATION SURVIVAL OF MURINE EMBRYOS VITRIFIED BY THE OPEN PULLED STRAW METHOD?

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab71 ◽

2011 ◽

Vol 23 (1) ◽

pp. 141

Author(s):

M. El-Gayar ◽

J. Reischl ◽

M. Gauly ◽

W. Holtz

Keyword(s):

Sucrose Solution ◽

Active Agent ◽

Virgin Female ◽

Control Group ◽

Embryo Survival ◽

Intact Mouse ◽

Group 4 ◽

Group 2

Vitrification of mammalian embryos comprises suspension in highly concentrated solutions of penetrating cryoprotectants. These cryoprotectants are known to be extremely cytotoxic to cells in an unfrozen state. In the present experiment, it was attempted to reduce the amount of cryoprotectants entering the cells to a minimum by at least partially dehydrating the blastomeres before exposing them to vitrification solutions. Sucrose is known to be a non-permeating osmotically active agent and is, therefore, suited to serve this purpose. It has been shown that concentrations of up to 1.1 M sucrose are well tolerated (Krag et al. 1985 Theriogenology 23, 199). Morphologically intact mouse blastocysts collected from superovulated 5- to 8-week-old virgin female NMRI mice were randomly allocated to 4 treatment groups (100 embryos/group). A control group (Group 1) was vitrified according to the slightly modified (El-Gayar et al. 2008 Cryobiology 57, 191–194) protocol of (Vajta et al. 1998 Mol. Reprod. Dev. 51, 53–58). The protocol involves exposure to 10% dimethyl-sulfoxide (Me2SO) + 10% ethylene glycol (EG) for 1 min; 20% Me2SO + 20% EG for 20 s; loading into straws and plunging directly into liquid nitrogen. In embryos of Group 2, the procedure mentioned above was preceded by exposure to a 1 M sucrose solution for 1 min. In Group 3, the procedure of Group 2 was followed; however, vitrification solution no. 1 (10% Me2SO + 10% EG) was omitted. In Group 4, sucrose was added to both vitrification solutions at a concentration of 0.2 mol with the 10% Me2SO + 10% EG-solution and of 0.4 mol with the 20% Me2SO + 20% EG-solution. After warming, embryos of all groups were cultured in vitro in microdrops of M16 medium for 48 h. Differences between means were tested for significance by the chi-square test. All embryos were recovered after warming. In the control group, 97% of the embryos had continued development to the expanded blastocyst stage and 89% had proceeded to hatch. The corresponding values for Groups 2, 3, and 4 were 81 and 64%; 60 and 29%, and 93 and 88%, respectively. Earlier studies have shown that in vitro hatching is closely correlated with in vivo survival (El-Gayar et al. 2008 Cryobiology 57, 191–194; Cryoletters 2010, in print). The differences between the control group and Groups 2 and 3 were significant (P < 0.01), whereas with Group 4 it was not (P > 0.05). Thus, exposure to sucrose before vitrification (Groups 2 and 3) compromised the viability of murine blastocysts rather than protecting them, whereas addition of sucrose to vitrification solutions did not. Because embryo survival after cryopreservation by the standard OPS procedure was exceptionally high, it was impossible to detect a potential improvement. This study was partly supported by a grant from the Egyptian government.

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Connecting METTL3 and intratumoural CD33+ MDSCs in predicting clinical outcome in cervical cancer

10.21203/rs.3.rs-27169/v2 ◽

2020 ◽

Author(s):

Huan-he Ni ◽

Lin Zhang ◽

He Huang ◽

Shu-qin Dai ◽

Jiang Li

Keyword(s):

Cervical Cancer ◽

Proportional Hazards ◽

Cox Model ◽

Disease Free Survival ◽

Suppressor Cells ◽

Cox Proportional Hazards ◽

Tumour Cells ◽

Rank Test ◽

Independent Factor

Abstract Background: Methyltransferase-like 3 (METTL3) is a member of the m6A methyltransferase family and acts as an oncogene in cancers. Recent studies suggest that host innate immunity is regulated by the enzymes controlling m6A epitranscriptomic changes. Here, we aim to explore the associations between the levels of METTL3 and CD33+ myeloid-derived suppressor cells (MDSCs) in tumour tissues and the survival of patients with cervical cancer (CC).Methods: Paraffin tumour specimens from 197 CC patients were collected. The expression levels of METTL3 and CD33 were measured by immunohistochemical (IHC) staining. The clinical associations of the IHC variants were analysed by Pearson's or Spearman's chi-square tests. Overall survival (OS) and disease-free survival (DFS) were estimated by the Kaplan-Meier method and log-rank test. Hazard ratios (HRs) and independent significance were obtained via Cox proportional hazards models for multivariate analyses. METTL3 in CD33+ cells or CC-derived cells was knocked down by METTL3-specific siRNA, and MDSC induction in vitro was performed in a co-culture system in the presence of METTL3-siRNA and METTL3-knockdown-CC-derived cells compared with that of the corresponding controls.Results: We found that tumour tissues displayed increased levels of METTL3 and CD33+ MDSCs compared with tumour-adjacent tissues from the same CC patients. Importantly, METTL3 expression was positively related to the density of CD33+ cells in tumour tissues (P = 0.011). We further found that the direct CD33+CD11b+HLA-DR- MDSC induction and tumour-derived MDSC induction in vitro were decreased in the absence of METTL3. The level of METTL3 in tumour microenvironments was significantly related to advanced tumour stage. The levels of METTL3 and CD33+ MDSCs in tumour tissues were notably associated with reduced DFS or OS. Cox model analysis revealed that the level of METTL3 in tumour cells was an independent factor for patient survival, specifically for DFS (HR = 3.157, P = 0.022) and OS (HR = 3.271, P = 0.012), while the CD33+ MDSC number was an independent predictor for DFS (HR: 3.958, P = 0.031). Interestingly, in patients with advanced-disease stages (II-IV), METTL3 in tumour cells was an independent factor for DFS (HR = 6.725, P= 0.010) and OS (HR = 5.140, P= 0.021), while CD33+ MDSC density was an independent factor for OS (HR = 8.802, P = 0.037).Conclusion: Our findings suggest that CD33+ MDSC expansion is linked to high levels of METTL3 and that METTL3 and CD33+ MDSCs are independent prognostic factors in CC.

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Connecting METTL3 and intratumoural CD33+ MDSCs in predicting clinical outcome in cervical cancer

10.21203/rs.3.rs-27169/v3 ◽

2020 ◽

Author(s):

Huan-he Ni ◽

Lin Zhang ◽

He Huang ◽

Shu-qin Dai ◽

Jiang Li

Keyword(s):

Cervical Cancer ◽

Proportional Hazards ◽

Cox Model ◽

Disease Free Survival ◽

Suppressor Cells ◽

Cox Proportional Hazards ◽

Tumour Cells ◽

Rank Test ◽

Independent Factor

Abstract Background: Methyltransferase-like 3 (METTL3) is a member of the m6A methyltransferase family and acts as an oncogene in cancers. Recent studies suggest that host innate immunity is regulated by the enzymes controlling m6A epitranscriptomic changes. Here, we aim to explore the associations between the levels of METTL3 and CD33+ myeloid-derived suppressor cells (MDSCs) in tumour tissues and the survival of patients with cervical cancer (CC).Methods: Specimens of paraffin embedded tumour from 197 CC patients were collected. The expression levels of METTL3 and CD33 were measured by immunohistochemical (IHC) staining. The clinical associations of the IHC variants were analysed by Pearson's or Spearman's chi-square tests. Overall survival (OS) and disease-free survival (DFS) were estimated by the Kaplan-Meier method and log-rank test. Hazard ratios (HRs) and independent significance were obtained via Cox proportional hazards models for multivariate analyses. METTL3 in CD33+ cells or CC-derived cells was knocked down by METTL3-specific siRNA, and MDSC induction in vitro was performed in a co-culture system in the presence of METTL3-siRNA and METTL3-knockdown-CC-derived cells compared with that of the corresponding controls.Results: We found that tumour tissues displayed increased levels of METTL3 and CD33+ MDSCs compared with tumour-adjacent tissues from the same CC patients. Importantly, METTL3 expression was positively related to the density of CD33+ cells in tumour tissues (P = 0.011). We further found that the direct CD33+CD11b+HLA-DR- MDSC induction and tumour-derived MDSC induction in vitro were decreased in the absence of METTL3. The level of METTL3 in tumour microenvironments was significantly related to advanced tumour stage. The levels of METTL3 and CD33+ MDSCs in tumour tissues were notably associated with reduced DFS or OS. Cox model analysis revealed that the level of METTL3 in tumour cells was an independent factor for patient survival, specifically for DFS (HR = 3.157, P = 0.022) and OS (HR = 3.271, P = 0.012), while the CD33+ MDSC number was an independent predictor for DFS (HR: 3.958, P = 0.031). Interestingly, in patients with advanced-disease stages (II-IV), METTL3 in tumour cells was an independent factor for DFS (HR = 6.725, P= 0.010) and OS (HR = 5.140, P= 0.021), while CD33+ MDSC density was an independent factor for OS (HR = 8.802, P = 0.037).Conclusion: Our findings suggest that CD33+ MDSC expansion is linked to high levels of METTL3 and that METTL3 and CD33+ MDSCs are independent prognostic factors in CC.

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In-vivo evaluation of the effect of cyanoacrylate on prosthetic vascular graft infection – does cyanoacrylate increase the severity of infection?

VASA ◽

10.1024/0301-1526/a000867 ◽

2020 ◽

Vol 49 (4) ◽

pp. 281-284

Author(s):

Atıf Yolgosteren ◽

Gencehan Kumtepe ◽

Melda Payaslioglu ◽

Cuneyt Ozakin

Keyword(s):

Vascular Graft ◽

Graft Infection ◽

Vascular Graft Infection ◽

Group 4 ◽

Significant Difference ◽

Group 3 ◽

Group 2 ◽

Prosthetic Vascular Graft Infection ◽

Group 1

Summary. Background: Prosthetic vascular graft infection (PVGI) is a complication with high mortality. Cyanoacrylate (CA) is an adhesive which has been used in a number of surgical procedures. In this in-vivo study, we aimed to evaluate the relationship between PVGI and CA. Materials and methods: Thirty-two rats were equally divided into four groups. Pouch was formed on back of rats until deep fascia. In group 1, vascular graft with polyethyleneterephthalate (PET) was placed into pouch. In group 2, MRSA strain with a density of 1 ml 0.5 MacFarland was injected into pouch. In group 3, 1 cm 2 vascular graft with PET piece was placed into pouch and MRSA strain with a density of 1 ml 0.5 MacFarland was injected. In group 4, 1 cm 2 vascular graft with PET piece impregnated with N-butyl cyanoacrylate-based adhesive was placed and MRSA strain with a density of 1 ml 0.5 MacFarland was injected. All rats were scarified in 96th hour, culture samples were taken where intervention was performed and were evaluated microbiologically. Bacteria reproducing in each group were numerically evaluated based on colony-forming unit (CFU/ml) and compared by taking their average. Results: MRSA reproduction of 0 CFU/ml in group 1, of 1410 CFU/ml in group 2, of 180 200 CFU/ml in group 3 and of 625 300 CFU/ml in group 4 was present. A statistically significant difference was present between group 1 and group 4 (p < 0.01), between group 2 and group 4 (p < 0.01), between group 3 and group 4 (p < 0.05). In terms of reproduction, no statistically significant difference was found in group 1, group 2, group 3 in themselves. Conclusions: We observed that the rate of infection increased in the cyanoacyrylate group where cyanoacrylate was used. We think that surgeon should be more careful in using CA in vascular surgery.

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