Mohammed F. Salama

Reversible drug-tolerant persister states are emerging as key drivers of limited therapeutic durability, offering a complementary non-genetic perspective distinct from traditional models of acquired resistance. This is of particular interest in lung adenocarcinoma where EGFR tyrosine kinase inhibitors (TKIs) elicit dramatic responses, yet residual surviving cells persist and ultimately seed relapse. To define mechanisms that enable survival during this earliest residual-disease phase, we focused on the drug-tolerant persister population that remains after EGFR TKI exposure and can later give rise to outgrowth. Initial observations of elevated transcript levels of PRKCA , which encodes PKCα, in established TKI-resistant models, together with markedly delayed tumor relapse following PKCα suppression in vivo, nominated PKCα as a candidate regulator of the persister-to-relapse transition. Genetic ablation of PRKCA or its inhibition with enzastaurin reduced residual survival and outgrowth after TKI exposure, indicating that PKCα functions as an early dependency of drug-tolerant persisters rather than as a general mediator of acquired resistance. Mechanistically, PKCα was required for persister-associated EMT, migratory capacity, and robust induction of ALDH1A1, the latter constraining oxidative stress and enhancing persister survival. Functionally, PKCα was specifically necessary for survival of a rare, pre-existing CD44 ^High stem-like subpopulation that exhibited marked plasticity and ultimately seeded persistence. Together, these data identify a PKCα-dependent EMT/stemness/ROS pathway as a critical survival program in EGFR TKI-tolerant persister cells and support therapeutic strategies aimed at eliminating residual disease to prolong clinical responses.

Objective: to evaluate the chemo -preventive effect of PPE on hepatic injury and hepatocarcinogenesis induced by DENA and CCL4 in male Wistar Rats. Design: Randomized controlled experimental study. Animals: Fifty male Wistar Rats . Procedures: Fifty male Wistar Rats were distributed equally into 5 groups. One group for HCC induction by DENA and CCL4, another 2 groups were pretreated with PPE 150 mg / kg /day and 250 mg / kg / day for two weeks before HCC induction. The other two groups were left as control groups, one received normal saline and the other one received PPE alone. At the end of the experiment, effect of PPE was evaluated . Results: Significant decrease of serum liver enzymes (ALT , AST , ALP ,GGT Total and Direct bilirubin ) and AFP were observed in HCC group pre treated with PPE with both doses at (p<0.05) . Significant increase in antioxidants levels (TAC ,CAT , SOD ,GPX ,GSH ) in liver tissue homogenate in HCC group pretreated with PPE with both doses . Also, histopathological examination of liver tissue in DENA /CCL4 treated group showed adenoma and carcinoma , but marked improving in hepatic tissue architecture and decreasing in score of adenoma and carcinoma in PPE treated group. Conclusion and clinical relevance: pomegranate phytochemicals extract ameliorates the hepatocarcinogenic effect induced by DENA and CCL4 by their anti-apoptotic effect, antioxidant effect that decrease ROS production and scavenge excess ROS present in tissue and decrease hepatotoxic effect of DENA and CCL4.

Objectives: The current work investigated the effect of Wharton jelly mesenchymal stem cells (WJ-MSCs) pretreated with all-trans-retinoic acid (ATRA) on renal ischemia in rats and the possible role of oxidative stress, apoptotic and Wnt/β-Catenin signaling pathways, and inflammatory cytokines in their effects. Methods: The study included 90 male Sprague Dawley rats that were allocated to five groups (n = 18 rats): (I) Sham-operated group (right nephrectomy was performed); (II) Ischemia/reperfusion injury (IRI) group, a sham group with 45-min renal ischemia on the left kidney; (III) ATRA group, an ischemic group with an intravenous (i.v.) administration of ATRA 10 µM, 10 min post-surgery); (IV) WJ-MSCs group, an IRI group with an i.v. administration of 150 µL containing 7 × 10⁶ WJ-MSCs, 10 min post-surgery; (V) WJ-MSCs + ATRA group, an IRI group with an i.v. administration of 150 µL of 7 × 10⁶ WJ-MSCs pretreated with 10 µM ATRA. At the end of the experiments, serum creatinine, BUN micro-albuminuria (MAU), urinary protein, markers of redox state in the left kidney (MDA, CAT, SOD, and GSH), and the expression of Bax, IL-6, HIF-1α, Wnt7B, and β-catenin genes at the level of mRNA as well as for immunohistochemistry for NFkB and β-Catenin markers were analyzed. Results: The current study found that 45-min of renal ischemia resulted in significant impairment of kidney function (evidenced by the increase in serum creatinine, BUN, and urinary proteins) and deterioration of the kidney morphology, which was associated with a significant increase in redox state (evidenced by an increase in MDA and a decrease in GSH, SOD, and CAT), and a significant increase in inflammatory and apoptotic processes (evidenced by an increase in Bax and IL-6, NFkB, Wnt7B, β-catenin and HIF-1α) in kidney tissues (p < 0.05). On the other hand, treatment with ATRA, WJ-MSCs, or a combination of both, caused significant improvement in kidney function and morphology, which was associated with significant attenuation of oxidative stress, apoptotic markers, and inflammatory cytokines (IL6 and NFkB) with the upregulation of HIF-1α and β-catenin in kidney tissues (p < 0.05). Moreover, the renoprotective effect of WJ-MSCs pretreated with ATRA was more potent than WJ-MSCs alone. Conclusions: It is concluded that preconditioning of WJ-MSCs with ATRA may enhance their renoprotective effect. This effect could be due to the upregulation of the beta-catenin/Wnt pathway and attenuation of apoptosis, inflammation, and oxidative stress.

Driver-directed therapeutics have revolutionized cancer treatment, presenting similar or better efficacy compared to traditional chemotherapy and substantially improving quality of life. Despite significant advances, targeted therapy is greatly limited by resistance acquisition, which emerges in nearly all patients receiving treatment. As a result, identifying the molecular modulators of resistance is of great interest. Recent work has implicated protein kinase C (PKC) isozymes as mediators of drug resistance in non-small cell lung cancer (NSCLC). Importantly, previous findings on PKC have implicated this family of enzymes in both tumor-promotive and tumor-suppressive biology in various tissues. Here, we review the biological role of PKC isozymes in NSCLC through extensive analysis of cell-line-based studies to better understand the rationale for PKC inhibition. PKC isoforms α, ε, η, ι, ζ upregulation has been reported in lung cancer, and overexpression correlates with worse prognosis in NSCLC patients. Most importantly, PKC isozymes have been established as mediators of resistance to tyrosine kinase inhibitors in NSCLC. Unfortunately, however, PKC-directed therapeutics have yielded unsatisfactory results, likely due to a lack of specific evaluation for PKC. To achieve satisfactory results in clinical trials, predictive biomarkers of PKC activity must be established and screened for prior to patient enrollment. Furthermore, tandem inhibition of PKC and molecular drivers may be a potential therapeutic strategy to prevent the emergence of resistance in NSCLC.

This study was carried out to investigate the potential effects of vitamin B12 and sitagliptin, and their possible synergistic effect with doxorubicin (DOX) on the Ehrlich solid tumor model. B12, sitagliptin, and their combination with DOX were administered to tumor‐bearing mice for 21 days. Treatment with B12, sitagliptin, as well as their combinations with DOX caused a significant inhibition of tumor growth and increased the survival time. Malondialdehyde levels and the relative expression of tumor necrosis factor‐α and nuclear factor kappa B were significantly decreased, whereas the total antioxidant capacity was significantly increased in all treated groups, except the DOX‐treated one, when compared with the positive control group. Moreover, increased apoptosis was also observed by increased cleaved caspase‐3 immunostaining and histopathological examination. In conclusion, the antitumor activity of B12 and sitagliptin could be attributed to their ability to induce apoptosis and suppress oxidative stress and inflammation.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Mutational activation of the epidermal growth factor receptor (EGFR) is a major player in the pathogenesis of non-small cell lung cancer (NSCLC). NSCLC patients with constitutively active EGFR mutations eventually develop drug resistance against EGFR tyrosine-kinase inhibitors; therefore, better understandings of key components of mutant EGFR (mtEGFR) signaling are required. Here, we initially observed aberrantly high expression of protein kinase Cα (PKCα) in lung adenocarcinomas, especially those with EGFR mutations, and proceeded to examine the role of PKCα in the regulation of the signaling pathways downstream of mtEGFR. The results showed that NSCLC cell lines with constitutively active EGFR mutations tend to have very or moderately high PKCα levels. Furthermore, PKCα was constitutively activated in HCC827 and H4006 cells which have an EGFR deletion mutation in exon 19. Interestingly, mtEGFR was not required for the induction of PKCα at protein and message levels suggesting that the increased levels of PKCα are due to independent selection. On the other hand, mtEGFR activity was required for robust activation of PKCα. Loss of functions studies revealed that the NSCLC cells rely heavily on PKCα for the activation of the mTORC1 signaling pathway. Unexpectedly, the results demonstrated that PKCα was required for activation of Akt upstream of mTOR but only in cells with the mtEGFR and with the increased expression of PKCα. Functionally, inhibition of PKCα in HCC827 led to caspase-3-dependent apoptosis and a significant decrease in cell survival in response to cellular stress induced by serum starvation. In summary, the results identified important roles of PKCα in regulating mTORC1 activity in lung cancer cells, whereby a primary switching occurs from PKCα-independent to PKCα-dependent signaling in the presence of EGFR mutations. The results present PKCα as a potential synergistic target of personalized treatment for NSCLC with constitutively active mutant forms of EGFR and constitutively active PKCα.

The objective of the present study was to evaluate the potential effect of dietary calcium butyrate on growth performance, carcass traits and gut health in Japanese quails. In total, 320 one‐day‐old Japanese quails were randomly assigned to 4 equal treatments, with 8 replicates of 10 Japanese quails, for 4 weeks. The Japanese quails in control treatment were fed control diet whereas in the other treatments the Japanese quails were fed diet supplemented with calcium butyrate at 0.3, 0.5 and 0.7 g/kg diet. Data concerning performance measurements were recorded weekly. In addition, eight Japanese quails (one/replicate) from each treatment were selected randomly for serum collection to measure pro‐ and anti‐inflammatory cytokines. Pooled faecal samples from each replicate of each treatment were also collected at three time points (0, 2 and 4 weeks) for count E. coli and C. perfringens. The results showed that after 7 days of the experimental period, Japanese quails fed calcium butyrate supplemented diet at 0.7 g/kg showed a greater (p < .05) body weight and a favourable (p < .05) feed conversion ratio than the other treatments. Moreover, serum superoxide dismutase and catalase activities were increased (p < .05) in Japanese quails fed calcium butyrate supplemented diet at 0.7 g/kg. Calcium butyrate supplementation at 0.7 g/kg was associated with reduction (p < .05) in TNF‐α, IL‐6 and IL1‐β, while IL‐10 was increased (p < .05). In addition, after 2 weeks of calcium butyrate supplementation, a reduction (p < .05) in E. coli and C. perfringens counts was observed in excreta of Japanese quails fed 0.5 and 0.7 g calcium butyrate/kg diets. It is concluded that calcium butyrate supplementation improves body weight gain, reduces E. coli and C. perfringens counts and has anti‐inflammatory/anti‐oxidant effect in Japanese quails.