作者: Metierre, Cynthia ; Nagarajah, Rajini ; Bailey, Charles G ; Yeo, Dannel ; Adelstein, Stephen ; Liang, Oliver ; Rasko, John E J ; Sagnella, Sharon M ; Castelletti, Laura ; Chu, Gerard J

BACKGROUND AIMSSeveral anti-mesothelin (MSLN) chimeric antigen receptor (CAR) T cells are in phase 1/2 clinical trials to treat solid-organ malignancies. The effect of MSLN antigen density on MSLN CAR cytotoxicity against tumor cells has not been examined previously, nor are there data regarding the effect of agents that increase MSLN antigen density on anti-MSLN CAR T cell efficacy.METHODSMSLN antigen density was measured on a panel of pancreatic cancer and mesothelioma cell lines by flow cytometry. In parallel, the cytotoxicity and specificity of two anti-MSLN CAR T cells (m912 and SS1) were compared against these cell lines using a real-time impedance-based assay. The effect of two MSLN 'sheddase' inhibitors (lanabecestat and TMI-1) that increase MSLN surface expression was also tested in combination with CAR T cells.RESULTSSS1 CAR T cells were more cytotoxic compared with m912 CAR T cells against cell lines that expressed fewer than ∼170 000 MSLN molecules/cell. A comparison of the m912 and amatuximab (humanized SS1) antibodies identified that amatuximab could detect and bind to lower levels of MSLN on pancreatic cancer and mesothelioma cell lines, suggesting that superior antibody/scFv affinity was the reason for the SS1 CAR's superior cytotoxicity. The cytotoxicity of m912 CAR T cells was improved in the presence of sheddase inhibitors, which increased MSLN antigen density.CONCLUSIONSThese data highlight the value of assessing CAR constructs against a panel of cells expressing varying degrees of target tumor antigen as occurs in human tumors. Furthermore, the problem of low antigen density may be overcome by concomitant administration of drugs that inhibit enzymatic shedding of MSLN.

2023-12-01·Journal of Molecular Medicine

Soluble LDL-receptor is induced by TNF-α and inhibits hepatocytic clearance of LDL-cholesterol

Article

作者: Zegeye, Mulugeta M ; Kumawat, Ashok K ; Nakka, Sravya S ; Sirsjö, Allan ; Andersson, Jonas S O ; Ljungberg, Liza U ; Söderberg, Stefan

AbstractDefective LDL-C clearance and hence its elevation in the circulation is an established risk factor for cardiovascular diseases (CVDs) such as myocardial infarction (MI). A soluble LDL-receptor (sLDL-R) has been detected in human plasma which correlates strongly with circulating LDL-C and classical conditions that promote chronic inflammation. However, the mechanistic interplay between sLDL-R, inflammation, and CVDs remains to be investigated. Here, we report that stimulation of HepG2 cells with TNF-α induces the release of sLDL-R into culture supernatants. In addition, TNF-α induces gene expression of peptidases ADAM-17 and MMP-14 in HepG2 cells, and inhibiting these peptidases using TMI 1 significantly reduces the TNF-α induced sLDL-R release. We found that a soluble form of recombinant LDL-R (100 nM) can strongly bind to LDL-C and form a stable complex (KD = E-12). Moreover, incubation of HepG2 cells with this recombinant LDL-R resulted in reduced LDL-C uptake in a dose-dependent manner. In a nested case-control study, we found that baseline sLDL-R in plasma is positively correlated with plasma total cholesterol level. Furthermore, a twofold increase in plasma sLDL-R was associated with a 55% increase in the risk of future MI [AOR = 1.55 (95% CI = 1.10–2.18)]. Nevertheless, mediation analyses revealed that a significant proportion of the association is mediated by elevation in plasma cholesterol level (indirect effect β = 0.21 (95% CI = 0.07–0.38). Collectively, our study shows that sLDL-R is induced by a pro-inflammatory cytokine TNF-α via membrane shedding. Furthermore, an increase in sLDL-R could inhibit hepatic clearance of LDL-C increasing its half-life in the circulation and contributing to the pathogenesis of MI.Key messages

TNF-α causes shedding of hepatocytic LDL-R through induction of ADAM-17 and MMP-14.

sLDL-R binds strongly to LDL-C and inhibits its uptake by hepatocytic cells.

Plasma sLDL-R is positively correlated with TNF-α and cholesterol.

Plasma sLDL-R is an independent predictor of myocardial infarction (MI).

Plasma cholesterol mediates the association between sLDL-R and MI.

2023-10-01·Poultry Science

Naked neck gene and intermittent thermal manipulations during embryogenesis improve posthatch performance and thermotolerance in slow-growing chickens under tropical climates

Article

作者: Bilalissi, Abidi ; Collin, Anne ; Dassidi, Nideou ; Assion, Mauril E P ; Meteyake, Hèzouwè T ; Tona, Kokou

Many studies have shown that thermal manipulations during the incubation (TMI) and naked neck gene (Na) positively affect heat-stressed broilers' thermotolerance, hatching process, and posthatch performance. Their combination could increase the beneficial effect on broilers reared under natural tropical climatic conditions. The aim of this study was to investigate the effects of the Na gene and TMI on hatching and posthatch performance of slow-growing broilers under tropical climates. The study included 1,200 hatching eggs from 2 different crosses: 1) females and males, both with a normal or fully feathered neck (na na group), and 2) females (with a normal neck) and males (bare neck) (Na na group), incubated in similar conditions until d 7. Thereafter, they were assigned to 3 subgroups for each cross: the control group (C) was incubated at standard incubation conditions (37.8°C, 60% RH). The TMI-1 group was subjected to TMI-1 (T = 38.5°C, RH = 65%, E10-18, 6 h/d) and TMI-2 group to TMI-2 (T = 39.5°C, RH = 65%, E7-16, 12 h/d). Between 450 and 504 h of incubation, eggs were checked for hatching events. During the posthatch phase, chicks from each incubation subgroups (Na na-C, Na na-TMI-1, Na na-TMI-2, na na-C, na na-TMI-1, na na-TMI-2) were raised for 12 wk at a tropical natural ambient temperature. Hatchability, hatching time, chick's temperature, final body weight (FBW), and feed conversion ratio (FCR) were determined. The results revealed that the Na gene reduced (P ˂ 0.05) hatchability. The control group had the highest mortality rate compared to TMI-1 and TMI-2 groups. There was an interaction between genotype and TMI on incubation duration, hatching weight, chick quality, FBW, and FCR (P ˂ 0.05). In conclusion, the Na gene influenced the effects of thermal manipulation. TMI-1 combined with Na gene improved the productive performances of broilers in a tropical climate.

100 项与 TMI-1 相关的药物交易

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