Combinatorial targeting of telomerase and DNA-PK induces synergistic apoptotic effects against Pre-B acute lymphoblastic leukemia cells.

BACKGROUND: Due to the high demand for novel approaches for leukemia-targeted therapy, this study investigates the impact of DNA-PK inhibitor NU7441 on the sensitivity of pre-B ALL cells to the telomerase inhibitor MST-312. METHODS: The study involved NALM-6 cells treated with MST-312 and NU7441, assessing their viability and metabolic activity using trypan blue and MTT assays. The study also evaluated apoptosis, gene expression changes, and DNA damage using flow cytometry, qRT-PCR, and micronucleus assays. The binding energy of MST-312 in the active site of telomerase was calculated using molecular docking. RESULTS: The study's findings revealed a synergistic decline in both cell viability and metabolic activity in NALM-6 cells when exposed to the combined treatment of MST-312 and NU7441, and this decrease occurred without any adverse effects on healthy PBMC cells. Furthermore, the combination treatment exhibited a significantly higher induction of apoptosis than treatment with MST-312 alone, as observed through flow cytometry assay. qRT-PCR analysis revealed that this enhanced apoptosis was associated with a notable downregulation of Bcl-2 expression and an upregulation of Bax gene expression. Moreover, the combination therapy decreased expression levels of hTERT and c-Myc genes. The micronucleus assay indicated that the combination treatment increased DNA damage in NALM-6 cells. Also, a good conformation between MST-312 and the active site of telomerase was revealed by docking data. CONCLUSIONS: The study suggests that simultaneous inhibition of telomerase and DNA-PK in pre-B ALL presents a novel targeted therapy approach.

Trioxidized cysteine in the aging proteome mimics the structural dynamics and interactome of phosphorylated serine.

Aging is the primary risk factor for the development of numerous human chronic diseases. On a molecular level, it significantly impacts the regulation of protein modifications, leading to the accumulation of degenerative protein modifications (DPMs) such as aberrant serine phosphorylation (p-Ser) and trioxidized cysteine (t-Cys) within the proteome. The altered p-Ser is linked to abnormal cell signaling, while the accumulation of t-Cys is associated with chronic diseases induced by oxidative stress. Despite this, the potential cross-effects and functional interplay between these two critical molecular factors of aging remain undisclosed. This study analyzes the aging proteome of wild-type C57BL/6NTac mice over 2 years using advanced proteomics and bioinformatics. Our objective is to provide a comprehensive analysis of how t-Cys affects cell signaling and protein structure in the aging process. The results obtained indicate that t-Cys residues accumulate in the aging proteome, interact with p-Ser interacting enzymes, as validated in vitro, and alter their structures similarly to p-Ser. These findings have significant implications for understanding the interplay of oxidative stress and phosphorylation in the aging process. Additionally, they open new venues for further research on the role(s) of these protein modifications in various human chronic diseases and aging, wherein exacerbated oxidation and aberrant phosphorylation are implicated.

Differential systemic decellularization in vivo to study molecular changes in each vasculature layer in murine models of disease.

Vascular dysfunction underlies the onset and progression of many life-threatening diseases, highlighting the need for improved understanding of its molecular basis. Here, we present differential systemic decellularization in vivo (DISDIVO), a protocol that enables systemic and independent study of the molecular changes in each vasculature layer in murine models of disease. We describe steps for anesthesia, perfusion surgery, and exsanguination. We then detail detachment and collection of glycocalyx and decellularization and collection of both endothelial and smooth muscle cells. For complete details on the use and execution of this protocol, please refer to Serra et al., Gallart-Palau et al., and Vinaiphat et al.1,2,3.

The Role of Genetics and Synergistic Effect of Targeting Common Genetic Mutations in Acute Lymphoblastic Leukemia (ALL).

Increasing concern regarding non-treatment and relapse in Acute Lymphoblastic Leukemia (ALL) among children and adults has attracted the attention of researchers to investigate the genetic factors of ALL and discover new treatments with a better prognosis. Nevertheless, the survival rate in children is more than in adults; therefore, it is necessary to find new potential molecular targets with better therapeutic results. Genomic analysis has enabled the detection of different genetic defects that are serious for driving leukemogenesis. The study of genetic translocation provides a better understanding of the function of genes involved in disease progression. This paper presents an overview of the main genetic translocations and dysregulations in the signaling pathways of ALL. We also report the inhibitors of these main translocations and evaluate the synergistic effect of chemical inhibitors and gamma-ray irradiation on ALL.

Inhibition of DNA damage response pathway using combination of DDR pathway inhibitors and radiation in treatment of acute lymphoblastic leukemia cells.

An alarming increase in acute lymphoblastic leukemia cases among children and adults has attracted the attention of researchers to discover new therapeutic strategies with a better prognosis. In cancer cells, the DNA damage response (DDR) pathway elements have been recognized to protect tumor cells from various stresses and cause tumor progression; targeting these DDR members is an attractive strategy for treatment of cancers. The inhibition of the DDR pathway in cancer cells for the treatment of cancers has recently been introduced. Hence, effective treatment strategies are needed for this purpose. Chemotherapy in combination with radiotherapy is considered a potential therapeutic strategy for acute leukemia. This review aims to assess the synergistic effects of these inhibitors with irradiation for the treatment of leukemia.

Lay abstract Acute lymphoblastic leukemia (ALL) is a blood malignancy that is caused by the high proliferation of lymphoid precursors in bone marrow and peripheral blood, and it is one of the common malignancies among children and adults. Novel therapeutic strategies are increasingly being studied to suppress cancer cells. DNA damage response pathways have developed in cancer cells that protect tumor cells from chemotherapy and radiotherapy and cause relapse and treatment failure. Effective drugs in ALL treatment can suppress cancer cells through several mechanisms, such as inducing DNA damage in cancer cells. However, due to the active repair systems in cancer cells, treatment may fail. Recently, the inhibition of these repair pathways and the combination of DNA damage response pathway inhibitors with chemotherapy and radiotherapy in cancer treatment have been studied. The purpose of this review is to outline the synergistic effects of DNA damage response pathway inhibitors and radiation in the death of leukemic cells.

Detection of Dermatophytes from Dermatophytosis-Suspected Cases in Iran, Evaluation of Polymerase Chain Reaction-Sequencing Method.

BACKGROUND: Dermatophytosis is mostly caused by dermatophytes species, and the diagnosis of disease is very important for early treatment. The aim of this study was to identify the commonly dermatophytes species isolated directly from the clinical samples, using the polymerase chain reaction (PCR) and evaluate both conventional and molecular methods. MATERIALS AND METHODS: This study was performed on 115 clinical samples. Dermatophyte isolates were initially identified by conventional method and confirmed by the sequencing molecular method. In this study, the molecular technique is implemented directly on clinical samples. Statistical analysis of the information was performed by the SPSS software, and the results were statistically analyzed. RESULTS: Our findings demonstrated that the most abundant dermatophyte species by PCR-sequencing were Trichophyton mentagrophytes (20%), followed by Trichophyton tonsurans (10%), Trichophyton rubrum (6.7%), T. interdigital (6.7%), Arthroderma otae, and Arthroderma vanbreuseghemii, (3.3%) for each one. CONCLUSION: For medical laboratories, routine procedures are still preferred because of their lower cost, and the results are almost the same as the molecular methods. The sensitivity and specificity values for PCR under our laboratory condition were 60% and 87%, respectively. This study shows that molecular results performed better in nails than other samples, by culture results.