Combination of p38 MAPK inhibitor with PD-L1 antibody effectively prolongs survivals of temozolomide-resistant glioma-bearing mice via reduction of infiltrating glioma-associated macrophages and PD-L1 expression on resident glioma-associated microglia

Background: Barretts Oesophagus (BO) presents a particular pathological dilemma, in that patients who have no dysplasia within their BO experience a small but significant risk of malignant progression each year. Screening programmes have attempted to reduce the mortality from BO associated oesophageal adenocarcinoma but cannot predict which BO patients will progress to invasive malignancy. We have previously identified the long non coding RNA, OR3A4, is differentially hypomethylated in progressive BO. We aimed to understand its role in BO pathogenicity Methods: The stable BO cell line CP-A, as well as the oesophageal adenocarcinoma cells line OE-33 was transfected with a lentiviral OR3A4 over-expression vector, and underwent high resolution microscopy, immunofluorescence, RT-qPCR, RNA sequencing, and targeted drug screening with the p38-MAPK inhibitor domipramod to understand the effects of OR3A4 expression on progression. We then compared progressive vs. non-progressive BO samples using quantitative multi-fluorophore (Vectra) immunohistochemistry. Results: Over-expression of OR3A4 in CP-A lines resulted in a hyperproliferative, dysplastic cellular phenotype, with strong over-expression of MAPK and anti-apoptotic pathways at the RNA and protein level, which was sensitive to the p38-MAPK inhibitor domipramod. Vectra immunohistochemistry demonstrated that progressive BO had reduced visibility associated with a reduction in CD8+ T-cells and CD68+ macrophages and reduced CD4+ T-cells in the stomal compartment. Conclusion: The overexpression of OR3A4, which we have previously shown is associated with progressive BO leads to a proliferative dysplastic cellular phenotype associated with increased, reversible MAPK signalling and loss of immune visibility.

Download Full-text

Role of Mitogen-Activated Protein Kinases in Activation of Human Neutrophils by Antineutrophil Cytoplasmic Antibodies

Journal of the American Society of Nephrology ◽

10.1681/asn.v12137 ◽

2001 ◽

Vol 12 (1) ◽

pp. 37-46

Author(s):

RALPH KETTRITZ ◽

ADRIAN SCHREIBER ◽

FRIEDRICH C. LUFT ◽

HERMANN HALLER

Keyword(s):

Flow Cytometry ◽

Cell Surface ◽

P38 Mapk ◽

Respiratory Burst ◽

Antineutrophil Cytoplasmic Antibodies ◽

Human Neutrophils ◽

Tnf Α ◽

Mitogen Activated Protein ◽

P38 Mapk Inhibitor ◽

Mapk Inhibitor

Abstract. Antineutrophil cytoplasmic antibodies (ANCA) may be important in the pathophysiology of necrotizing vasculitis. ANCA activate human neutrophils primed with tumor necrosis factor-α (TNF-α) in vitro. TNF-α priming results in translocation of ANCA antigens to the cell surface, where they are recognized by the antibodies. The signaling mechanisms involved in TNF-α priming and subsequent ANCA-induced activation were investigated. TNF-α-primed neutrophils were stimulated with monoclonal antibodies (MAb) to human myeloperoxidase (MPO) and proteinase 3 (PR3), and with preparations of human ANCA (three patients with PR3-ANCA and two patients with MPO-ANCA). Respiratory burst was measured with superoxide dismutase-inhibitable ferricytochrome C reduction and using dihydro-rhodamine-1,2,3. Phosphorylation of p38 mitogen-activated protein kinase (p38-MAPK) and the extracellular signal-regulated kinase (ERK) were assessed by immunoblotting. ANCA-antigen translocation was studied by flow cytometry. The tyrosine phosphorylation inhibitor genistein, but not calphostin or staurosporin, resulted in a significant dose-dependent superoxide generation inhibition (11.6 ± 1.7 nmol to 2.1 ± 0.5 for PR3-ANCA, and 16.0 ± 2.8 to 3.3 ± 1.3 for MPO-ANCA). The p38-MAPK inhibitor (SB202190) and the ERK inhibitor (PD98059) diminished PR3-ANCA-mediated superoxide production dose dependently (11.6 ± 1.7 nmol O2- to 1.9 ± 0.6 with 50 μM SB202190 and 4.0 ± 0.6 with 50 μM PD098059, respectively). For MPO-ANCA, the results were similar (16.0 ± 2.8 nmol to 0.9 ± 1.0 nmol with SB202190 and 6.4 ± 2.4 nmol with PD98059, respectively). Western blot showed phosphorylation of both p38-MAPK and ERK during TNF-α priming. The p38-MAPK inhibitor and the ERK inhibitor showed the strongest effect on respiratory burst when added before TNF-α priming, further supporting an important role for both signaling pathways in the priming process. Flow cytometry showed that p38-MAPK inhibition decreased the translocation of PR3 (by 93 ± 2%) and of MPO (by 64 ± 2%). In contrast, no such effect was seen when ERK was inhibited. Thus, p38-MAPK and ERK are important for the TNF-α-mediated priming of neutrophils enabling subsequent ANCA-induced respiratory burst. However, both pathways show differential effects, whereby p38-MAPK controls the translocation of ANCA antigens to the cell surface.

Download Full-text

Effects of p38 MAPK inhibitors on fibroblast differentiation in the zone of postoperative scar formation

Nauchno-prakticheskii zhurnal «Patogenez» ◽

10.25557/2310-0435.2018.02.43-47 ◽

2018 ◽

pp. 43-47

Author(s):

Ирина Александровна Шурыгина ◽

Н.В. Зеленин ◽

Г.Б. Гранина ◽

М.Г. Шурыгин

Keyword(s):

P38 Mapk ◽

Wistar Rats ◽

Scar Formation ◽

Control Group ◽

P38 Inhibitor ◽

P38 Mapk Inhibitors ◽

Mapk Inhibitors ◽

P38 Mapk Inhibitor ◽

Postoperative Scar ◽

Mapk Inhibitor

Цель исследования: оценить влияние блокатора р38 МАРК SB203580 на дифференцировку фибробластов в зоне формирования послеоперационного рубца. Материалы и методы: На модели линейной кожно-мышечной раны у крыс линии Вистар оценено влияние локального введения блокатора р38 МАРК SB203580 в составе лекарственной пленки на дифференцировку фибробластов (n = 30) по сравнению с заживлением раны без введения активного вещества (n = 30). Проводили оценку количества коллагена в области раны, периоперационной зоне и интактной дерме, а также иммуноморфологическое окрашивание на CD34, CD45, ММР9 и актин в сроки от 2 часов до 30 суток. Результаты: Установлено, что применение блокатора р38 SB203580 приводило к значительному снижению интенсивности коллагенообразования в зоне формирующегося рубца. Так, у животных контрольной группы относительная площадь, занятая волокнами коллагена в зоне послеоперационной раны, закономерно возрастала в сроки от 3 до 30 суток, достигая максимальных значений к концу наблюдения - 73,54% [66,87; 78,01]. При введении SB203580 в течение всего срока наблюдения отмечалось достоверное снижение коллагенообразования в сравнении с группой контроля, к 30 суткам показатель составил 43,60% [41,05; 60,15] (р = 0.002). Введение SB203580 снижало привлечение прогениторных клеток фибробластического ряда в зону формирования послеоперационного рубца, повышало фиброкластическую активность. Aim. To assess effects of a p38 MAPK inhibitor, SB203580, on fibroblast differentiation in the zone of postoperative scar formation. Materials and methods. The effect of locally injected p38 MAPK inhibitor, SB203580, on fibroblast differentiation (n = 30) was compared with wound healing without an active substance injection (n = 30) on a model of linear musculocutaneous wound in Wistar rats weighing 220-250 g. We measured the amount of collagen in the wound zone, perioperative zone, and intact derma and conducted immunomorphological staining for CD34, CD45, MMP9, and actin at timepoints of 2 hours to 30 days. Results. The p38 inhibitor, SB203580, induced a significant decrease in collagenation intensity in the zone of forming scar. In Wistar rats of the control group, the percent area of collagen fibers in the zone of postoperative wound was increasing between days 3 and 30 and reached a maximum of 73.54 % [66.87; 78.01] by the end of observation period. The SB203580 injection significantly decreased collagenation over the entire period of observation compared with the control group (43.60 % [41.05; 60.15] by day 30, р = 0.002). The SB203580 injection decreased the engagement of fibroblastic progenitors in the zone of postoperative scar formation and increased the fibroclast activity.

Download Full-text

P38 MAPK Pharmacological Inhibitor SB203580 Alleviates Total Parenteral Nutrition-Induced Loss of Intestinal Barrier Function but Promotes Hepatocyte Lipoapoptosis

Cellular Physiology and Biochemistry ◽

10.1159/000457933 ◽

2017 ◽

Vol 41 (2) ◽

pp. 623-634 ◽

Cited By ~ 9

Author(s):

Yong-Tao Xiao ◽

Wei-Hui Yan ◽

Yi Cao ◽

Jun-Kai Yan ◽

Wei Cai

Keyword(s):

Parenteral Nutrition ◽

Palmitic Acid ◽

Total Parenteral Nutrition ◽

P38 Mapk ◽

Barrier Function ◽

Intestinal Barrier ◽

Mitogen Activated Protein Kinase ◽

Intestinal Barrier Function ◽

P38 Mapk Inhibitor ◽

Mapk Inhibitor

Background & Aims: Our previous studies have provided evidence that p38 mitogen-activated protein kinase (MAPK) is involved in total parenteral nutrition (TPN)-associated complications, but its exact effects and mechanisms have not been fully understood. This study aimed to evaluate the roles of p38 MAPK inhibitor SB203580 in the TPN-induced loss of intestinal barrier function and liver disease. Methods: A rodent model of TPN was used to analyze the roles of SB203580 in TPN-associated complications.Intestinal barrier function was evaluated by transepithelial electrical resistance (TER) and paracellular permeability in Caco-2 cells. The palmitic acid (PA) was used to induce hepatic lipoapoptosis in vitro. The lipoapoptosis was detected using Caspase-3/7 and lipid staining. Results: In the present study, we showed that SB203580 treatment significantly suppressed TPN-mediated intestinal permeability in rats. SB203580 treatment significantly inhibited IL-1β-induced an increase in tight junction permeability of Caco-2 cells via repressing the p38/ATF-2 signaling. Unexpectedly, SB203580 treatment enhanced hepatic lipoapoptosis in the model of TPN. Palmitic acid (PA)-induced hepatic lipoapoptosis in human liver cells was significantly augmented by the SB203580 treatment. Conclusions: We demonstrate that the p38 MAPK inhibitor SB203508 ameliorates intestinal barrier function but promotes hepatic lipoapoptosis in model of TPN.

Download Full-text