Activation Of Proto-oncogenes Research Articles

Roles of oncogenes in esophageal squamous cell carcinoma and their therapeutic potentials.

Esophageal squamous cell carcinoma (ESCC) is the most common type of esophageal cancer (EC) in Asia. It is a malignant digestive tract tumor with abundant gene mutations. Due to the lack of specific diagnostic markers and early cancer screening markers, most patients are diagnosed at an advanced stage. Genetic and epigenetic changes are closely related to the occurrence and development of ESCC. Here, We review the activation of proto-oncogenes into oncogenes through gene mutation and gene amplification in ESCC from a genetic and epigenetic genome perspective, We also discuss the specific regulatory mechanisms through which these oncogenes mainly affect the biological function and occurrence and development of ESCC through specific regulatory mechanisms. In addition, we summarize the clinical application value of these oncogenes is summarized, and it provides a feasible direction for clinical use as potential therapeutic and diagnostic markers.

Research on the influence of APOBEC family on the occurrence, diagnosis, and treatment of various tumors.

Apolipoprotein B mRNA-editing catalytic polypeptide (APOBEC) is a family of highly efficient cytidine deaminase enzymes. APOBECs have been proven to deaminate cytidine on single-stranded DNA or RNA. Inducing the deamination of cytosine on the target gene into uracil, which exerts a variety of physiological functions, plays an important role in innate immunity, adaptive immunity, and antiviral. As the research progresses, APOBECs have been confirmed to be highly expressed in a variety of tumors, causing abnormal mutations in host genes, leading to inactivation of tumor suppressor genes or activation of proto-oncogenes, and their role in tumor development and as diagnostic and treatment markers gradually be found. This article will review the mechanism of APOBECs and their impact on tumor occurrence, development, diagnosis, and treatment, and provide a theoretical basis for future tumor treatment.

Precision oncology for RET-related tumors.

Aberrant activation of the RET proto-oncogene is implicated in a plethora of cancers. RET gain-of-function point mutations are driver events in multiple endocrine neoplasia 2 (MEN2) syndrome and in sporadic medullary thyroid cancer, while RET rearrangements are driver events in several non-medullary thyroid cancers. Drugs able to inhibit RET have been used to treat RET-mutated cancers. Multikinase inhibitors were initially used, though they showed modest efficacy and significant toxicity. However, new RET selective inhibitors, such as selpercatinib and pralsetinib, have recently been tested and have shown good efficacy and tolerability, even if no direct comparison is yet available between multikinase and selective inhibitors. The advent of high-throughput technology has identified cancers with rare RET alterations beyond point mutations and fusions, including RET deletions, raising questions about whether these alterations have a functional effect and can be targeted by RET inhibitors. In this mini review, we focus on tumors with RET deletions, including deletions/insertions (indels), and their response to RET inhibitors.

Quantum technology a tool for sequencing of the ratio DSS/DNA modifications for the development of new DNA-binding proteins

ABSTRACT The current study summarizes the literature on quantum-based structural sequencing methods and their predicted roles in the discovery of new DNA-binding protein domains. Although there is no literature yet showing the role of quantum structural sequencing for the discovery of new domains for DNA-binding proteins, the current study shows that the development of new quantum-based and far-from-equilibrium biological systems can be achieved by 3D-curved room-temperature superconductivity materials. The technology is useful for protein engineering and 3D sequencing of the ratio DNA/RNA secondary structures (DSS)/Physiological Modifications (PM) and will overcome the use of nanopore and ChiP sequencing. This approach is very useful for the protein engineering of recombinant enzymes for the unnatural replication of proteins and glycans for new biochemical activities, treatment of HIV and cancers, new genome editing machinery, artificial replication and translation, first sequencing of DSS/physiological modification/telomere with a correlation to cancer genesis and therapy, The key DSS in this study are: quadruplex, hairpin loop, triplexes and duplexes, i-motif, Holliday junction; and the PM investigated: 6 mA, 6fmA, 5mC, 5fC, 8-oxo-guanine. The DSS/PM ratio is critical for proto-oncogene activation and cancer genesis.

Nicotinamide N-methyltransferase expression in squamous cell carcinoma of the vulva

The molecular pathways involved in the development of vulvar squamous cell carcinoma (SCC) cancer are not completely known. Nicotinamide N-methyltransferase (NNMT) is a cytosolic enzyme associated with tumorigenesis and metastasis in a variety of cancers. Its role in vulvar cancer has not been studied, previously. Vulvar SCC, high and low grade squamous intraepithelial lesions (SILs) and benign squamous hyperplasia were analysed immunohistochemically. The mean staining score for vulvar SCC was significantly higher than the score for vulvar squamous hyperplasia (p<.001). The mean relapse-free survival for patients with low and high NNMT expression was 41.4 months (95% CI: 25.6–57.2) and 19.8 months (95% CI: 3.0–36.6), respectively (p=.035). The mean disease-specific survival for patients with low and high NNMT expression was 75.8 months (95% CI: 57.5–94.2) and 27.8 months (95% CI 12.2–43.4), respectively (p=.015). Although quite preliminary, this study showed that NNMT expression was elevated in vulvar SCC compared to benign and premalignant lesions. Additionally, elevated NNMT expression was associated with poor survival. Impact Statement What is already known on this subject? Nicotinamide N-methyltransferase (NNMT) is a methyltransferase, associated with tumour progression, spread and poor prognosis in a variety of cancers. Its upregulation can lead to DNA hypomethylation, which can in turn result in the activation of proto-oncogenes and deactivation of tumour suppressor genes. What do the results of this study add? Although quite preliminary, this study showed that NNMT expression was elevated in vulvar SCC compared to benign and premalignant lesions. Additionally, elevated NNMT expression was associated with poor survival. What are the implications of these findings for clinical practice and/or further research? NNMT has been regarded as a potential target of cancer therapy and its role in vulvar cancer has not been studied, previously. This is the first study to investigate the expression of NNMT in vulvar cancer and associate NNMT elevation with poor survival. NNMT can further be investigated as a possible target of vulvar cancer therapy.

Pyroptosis- What We Know and the Road Ahead!

Pyroptosis, a type of cell death, initiated by proinflammatory signals, is classically associated with inflammation. It was thought to occur mainly in macrophages and leukocytes during inflammatory conditions and involves the activation of caspase 1 {Interleukin1beta (IL-1β)-converting enzyme} followed by the production of proinflammatory cytokines such as IL-1β. The process takes place within dendritic cells and macrophages, although there have been few reports of caspase-1 activity within other cells. Induction of pyroptosis involves the caspase-1 activity, however, how its downstream substrates bring about cell death is not well understood. And recently, there has been an addition to the gasdermin family, namely, Gasdermin-D (GSDMD) protein, which plays an important role during pyroptosis. Pyroptosis has been found to have a significant role in tumourigenesis, as it extrudes inflammatory mediators which are vital for tumour development. It is also well known that the activity of proto-oncogenes and anti-oncogenes, the microenvironment of the immune system, chronic inflammation, and oxidative stress, could determine tumourigenesis and hence the link to pyroptosis.

An Observational Cross-sectional Study Section of Factors Causing Delay in Diagnosis of Head and Neck Cancers at a Rural Tertiary Care Centre in Northern India

Introduction: Cancer is a complex genetic disease derived from the accumulation of a variety of genetic changes, which include activation of proto-oncogenes and inactivation of tumour suppressor genes. Head and Neck Cancers (HNC) are rapidly growing tumours with a median potential doubling time of only 6-7 days. Although these cancers are easily accessible, patients rarely present early. In India, most HNCs present in advanced stages resulting in increased morbidity and mortality. Hence, it is prudent to ascertain the factors which lead to delay in diagnosis of HNC as early diagnosis and treatment is the cornerstone in reducing consequences of HNC. Aim: To identify patient and professional factors causing delay in diagnosis of HNC and to identify association if any with stage of cancer presentation. Materials and Methods: An observational cross-sectional study was conducted in the Department of Ear Nose and Throat, at BPS Government Medical College for women, Sonepat, Haryana, India, from September 2019 to January 2021. Study included 55 newly diagnosed histopathologically confirmed head and neck cancer patients. Clinico-demographic details were inferred using elaborate clinical examination. Data were described in terms of range, mean±standard deviation, frequency and relative frequency (percentages). Chi-square and Fisher’s-exact test was used for comparing categorical data. Delay due to patient and professional factors were calculated separately and the association of each delay with important variables were compared. Results: Most patients (43.6%) belonged to the age group of 51 to 60 years, and the mean age was 60.25±9.81 years. Majority of the patients were males (85.5%). The mean total delay from onset of symptoms to final diagnosis was 22.38±7.23 weeks. Delayed patient presentation was the main cause of total delay in diagnosis. The main factors responsible for patient delay were rural residential status, low educational, socio-economic status and poor cancer awareness. Irrational therapies still contributed significantly for delayed patient presentation. Diagnostic delay led to upstaging of disease. Conclusion: Patient delay is the main factor responsible for delay in diagnosis of HNC. Even in this era of easy availability and accessibility of information, lack of awareness still exists at the fundamental level.

Mapping Scientific and Technological Production Related to the MYC Gene

In appropriate activation of c-MYC proto-oncogene contributes to the development of human cancers. Searches for therapies that target genes and proteins related to neoplastic phenotypes have become frequent. Therefore, inhibiting c-MYC expression has been the target for developing and testing multiple drugs and strategies for the treatment of various human cancers. This study aimed to map scientific and technological productions on the MYC gene at the Scielo, PubMed and Orbit Intelligence platforms between 2000 and 2019. The scientific prospecting revealed 1,259 articles. The most detected categories were: molecular biology, MYC mutations and those addressing the MYC as a drug target or therapeutic strategies. A progressive increase in the number of articles in this last category was found. Technological mapping detected 10,059 patent documents, with 20.2% granted. China and the USA were the largest filers, accounting for more than 40%. Biotechnology was the field with the highest number of patents. Biotechnology and the pharmaceutical sector predominated in the second half of the period investigated, both in scientific and technological prospecting. Our study points to a scientific and technological effort in the development of therapeutic strategies against cancer, in which MYC is among the main targets.

RBM15 promotes hepatocellular carcinoma progression by regulating N6-methyladenosine modification of YES1 mRNA in an IGF2BP1-dependent manner

The function of the N6-methyladenosine (m6A) methyltransferase RNA-binding motif protein 15 (RBM15) in hepatocellular carcinoma (HCC) has not been thoroughly investigated. Here we determined the clinical value, biological functions, and potential mechanisms of RBM15 in HCC. Expression of RBM15 was identified using tissue microarrays and online databases. A risk-prediction model based on RBM15 was developed and validated. We determined the biological role of RBM15 on HCC cells in vitro and in vivo. RNA sequencing was used to screen candidate targets of RBM15. Subsequently, the m6A dot blot assay, methylated RNA immunoprecipitation qPCR, dual-luciferase reporter assays, RNA decay assay, and RNA immunoprecipitation qPCR were employed to explore the mechanisms of RBM15. Our study showed that RBM15 was highly expressed in HCC, and overexpression of RBM15 indicated a worse outcome. A new nomogram combining RBM15 with age and TNM stage was developed and validated to predict the outcome of HCC patients; our nomogram increased the prediction accuracy of the TNM system. Functionally, RBM15 facilitates the proliferation and invasiveness of HCC. RBM15-mediated m6A modification contributed to a post-transcriptional activation of YES proto-oncogene 1 (YES1) in an insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1)-dependent manner. In addition, YES1 was confirmed as an oncogene in HCC cells by activating the mitogen-activated protein kinase (MAPK) pathway. In conclusion, RBM15-mediated m6A modification might facilitate the progression of HCC via the IGF2BP1–YES1–MAPK axis. RBM15 may be a promising biomarker in the outcome prediction of HCC.

The effect of survivin gene in breast cancer risk and prognosis

Abstract Objectives The accumulation of genetic damages in onset of cancer induce activation of protooncogenes or inactivation of tumor suppressor genes thus cause disruption of the balance between cell proliferation and programmed cell death. As a member of the apoptosis inhibitory protein family (IAP), survivin play important roles in carcinogenesis process. The evidence suggests that polymorphisms located in survivin promoter region may be important in determining genetic susceptibility of cancer. In this study, we aimed to examine a possible role of survivin −31 and −625 G/C gene polymorphisms in breast cancer. Methods A total of 160 breast cancer cases and 153 healthy controls were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methodology. Results Genotype and allele distributions and of −31 and −625 G/C polymorphisms were not significantly different between two groups. However, we observed the carriers of survivin −625 C/G polymorphism homozygous genotypes (GG/CC) were the significantly higher in patients with tumor necrosis (p=0.047). Conclusions Our results suggest that survivin −625 C/G polymorphism may be related with tumor prognosis, but we are opinion of that our result require to be validated in larger samples and further comprehensive research may explore the correlation.

Immunohistochemical evaluation of survivin in oral lichen planus and oral squamous cell carcinoma - a retrospective study.

Oral cancer is a multistep process involving enhanced activation of proto-oncogenes. Survivin (anti-apoptotic protein) is one of such proto-oncogenes that contribute to the carcinogenesis by deregulating cell proliferation. Expression of these proteins in cancerous and pre-cancerous lesions helps a better understanding of the etiology, treatment and prognosis. To evaluate qualitative and quantitative expression of survivin in healthy mucosa, oral lichen planus (OLP) and oral squamous cell carcinoma (OSCC). A total of 70 cases of formalin fixed paraffin embedded blocks, of which 20 healthy mucosa, 20 OSCC and 30 OLP were included in the study and stained immunohistochemically using antibodies against survivin monoclonal antibody. 4 μm sections were stained immunohistochemically, both nuclear and cytoplasmic staining was considered as positive for surviving. Positive and negative stained cells were counted with the help of Olympus BX51 rarefaction microscope and were analyzed quantitatively using image analysis pro plus software. Representative fields were randomly selected at 40× high power magnification. Values obtained were evaluated and tabulated for statistical analysis. There was a statistically significant relation between the expressions of survivin among healthy mucosa, OLP, OSCC with a p value of 0.001. Presence of survivin expression in healthy mucosa, OLP and OSCC, which is an inhibitor of apoptosis protein, can be identified as a useful diagnostic tool for the identification of precancerous lesions and conditions which are at higher risk for progression into invasive carcinoma.

Genetic Activation and Therapeutic Targeting of PIM1 in T-Cell Acute Lymphoblastic Leukemia/Lymphoma

T-cell acute lymphoblastic leukemia (T-ALL) and T-cell acute lymphoblastic lymphoma (T-LBL) are aggressive immature T-cell malignancies that are often characterized by improper T-cell receptor (TCR) recombinations leading to aberrant activation of proto-oncogenes. Current multi-agent combination chemotherapy treatment provides an overall survival rate of ~80% in children and 50% in adults. However, the prognosis of patients with relapsed and refractory T-ALL/T-LBL remains extremely poor and the development of more effective and specific antileukemic drugs is critical. Therefore, we aim to identify new oncogenes, define their mechanism of action and establish strategies to develop targeted, highly specific antileukemic drugs.Here, we used Targeted Locus Amplification (TLA, de Vree et al., Nat Biotechnol., 2014) to identify a novel TCRβ driven t(6;7)(p21;q34) translocation in a human T-LBL patient resulting in aberrant activation of the PIM1 proto-oncogene. PIM1 is a constitutively active serine/threonine kinase involved in cell cycle progression, apoptosis, transcription and drug resistance and is overexpressed in a variety of human cancers. Its expression is controlled by the JAK/STAT signaling pathway. Further characterization of this PIM1 rearranged patient sample revealed cooperative genetic alterations that target known T-ALL/T-LBL onco- and tumor suppressor genes, including NOTCH1, IKZF1, EP300 and CDKN2A.Comparing PIM1 expression between normal T-cell subsets, T-ALL and T-LBL patient samples using qPCR revealed subsets in both T-LBL and T-ALL patients with high PIM1 levels. This PIM1 expression can be both monoallelicaly (due to translocation) or biallelicaly (due to upstream mutations in f.e. the JAK/STAT pathway). Using publically available gene expression datasets, we show that oncogenic PIM1 activation is largely restricted to immature, TLX1, TLX3, NKX2-1 or HOXA positive T-ALL and T-LBL patients, and is mostly not present in TAL1 or LMO2 rearranged tumors. This pattern of PIM1 activation is in line with the preferential occurrence of JAK-STAT pathway alterations in these genetic subtypes of human T-ALL/T-LBL.To study the oncogenic properties of PIM1 in the context of malignant T-cell transformation, we did RNA sequencing and phosphoproteomics on the T-ALL/T-LBL tumor line HSB-2 (cell line with the highest PIM1 expression) and on ex vivo treated leukemic cells harvested from the spleen of T-LBL engrafted NSG mice after PIM1 inhibition with TP-3654 (Foulks et al., Neoplasia, 2014). These data revealed that PIM1 inhibition has broad effects on transcription and phosphorylation substrates involved in cell cycle, translation and apoptosis. For example, we showed that the oncogenic properties of PIM1 are, at least in part, mediated by phosphorylation of GSK3β and subsequent stabilization of anti-apoptotic MCL1.Next, we evaluated the therapeutic potential of PIM1 inhibition. Daily TP-3654 treatment for 4 weeks of T-LBL engrafted NSG mice resulted in strong anti-leukemic effects. In vitro combination treatment with TP-3654 and glucocorticoids showed strong synergism, while the combination with other chemotherapeutics such as asparaginase and vincristine did not. Moreover, the combination of TP-3654 and the glucocorticoid dexamethasone in vivo almost completely cured the leukemic mice (<1% hCD45 positive blasts in the peripheral blood).All together, our study identifies PIM1 as a putative oncogene in T-ALL and T-LBL and suggests that inhibition of this serine/threonine kinase could serve as a novel therapeutic strategy in these aggressive T-cell neoplasms. DisclosuresNo relevant conflicts of interest to declare.

Prognostic factors in RET dependent cancers treateded with RET inhibitors in early phase clinical trials.

3117 Background: Activation of the RET proto-oncogene has been identified in multiple cancer types, for example, gene rearrangements in non-small cell lung cancer (NSCLC) and papillary thyroid cancer (PTC) and activating mutations in medullary thyroid cancer (MTC), amongst others. The recent FDA approval of two highly selective RET inhibitors, selpercatinib and pralsetinib has changed the treatment paradigm of RET-driven cancers, but the significance of historical prognostic factors is unknown. Herein, we analyzed the outcomes of patients with RET-altered cancers enrolled in phase I trials and assess the utility of prognostic scores. Methods: A retrospective analysis was performed of patients treated with selective RET inhibitors at the MD Anderson Cancer Center (MDACC). Baseline clinical factors and survival data were assessed. Overall and progression free survival (OS and PFS) were estimated using the Kaplan-Meier method and multivariable Cox models were constructed. For all a p-value of &lt; 0.05 was consider significant. Results: Among 126 patients, median age was 58 years (range, 15-82), most with ECOG 0-1 (n = 124). RET-mutant MTC was most frequent (n = 81), followed by RET fusion-positive NSCLC (n = 30) and RET fusion positive thyroid cancer (n = 9). RET fusion partners were KIF5B (n = 17), CCD6 (n = 12) and NCOA4 (n = 7). RET M918T mutation was the most frequent (n = 50, 63%). Most patients were treated in the relapsed/refractory (R/R) setting (n = 85) and received a median of 1 prior line of therapy (range, 0-11). Median follow up was 20 months (range, 1-43). The estimated median PFS and OS were 24 and 35 months, respectively. Overall objective response rate was 64% (81/126), 2 complete response, 79 partial response, 32 had stable disease (25%) and 13 had progressive disease (PD). The following were associated with shorter PFS and OS: age ≥50 years (p &lt; 0.05), albumin &lt; 4 g/dL (p &lt; 0.01), brain metastases (p &lt; 0.0001), hemoglobin &lt; 12 g/dL ( &lt; 0.05), LDH &gt; normal (p &lt; 0.05), WBC ≥8 (p &lt; 0.01), PD (p &lt; 0.0001) and NSCLC (p &lt; 0.01). The M918T mutation and ECOG &gt; 0 were associated with shorter OS but not PFS (p &lt; 0.05). &gt; 3 metastatic sites and R/R disease were associated with inferior PFS (p = 0.04 and p = 0.01, respectively) but not OS. The Royal Marsden Hospital (RMH) and MDACC prognostic scores were significantly associated with PFS and OS (p &lt; 0.01). In multivariable models including all variables significantly associated with PFS and OS (excluding LDH as this was only tested in 58 patients) albumin &lt; 4 (HR 6.10, p = 0.013), brain metastases (HR 4.90, p = 0.027) and WBC ≥8 (HR 4.67, p = 0.031) were associated with inferior OS. NSCLC was significantly associated with inferior PFS (HR 5.45, p = 0.02). Conclusions: The RMH and MDACC prognostic scores predict OS in RET-aberrant cancers treated on early phase trials. Low albumin, WBC &gt; 8 and brain metastases are significantly associated with inferior survival.

The Role of p53 Dysfunction in Colorectal Cancer and Its Implication for Therapy.

Simple SummaryDespite remarkable progress being made by preventive medical check-ups in the last decades, colorectal cancer (CRC) remains one of the most frequent and deadliest cancers worldwide. An understanding of the mutational landscape in CRC is needed to develop new mutanome-directed therapies with stronger efficacy and less side-effects than current therapeutic standard regimes. Carcinogenesis in CRC is a multi-gene driven process, where premalignant cells accumulate successively key tumorigenesis-related mutations. Here, inactivation of the tumor suppressor gene p53 is a hallmark event during tumorigenesis. Mutations of p53 impact the prognosis of patients, enabling the use of targeted therapies such as immune therapy. Alterations of p53 affect not only the tumor biology of cancer cells but also the surrounding tumor microenvironment (TME).Colorectal cancer (CRC) is one of the most common and fatal cancers worldwide. The carcinogenesis of CRC is based on a stepwise accumulation of mutations, leading either to an activation of oncogenes or a deactivation of suppressor genes. The loss of genetic stability triggers activation of proto-oncogenes (e.g., KRAS) and inactivation of tumor suppression genes, namely TP53 and APC, which together drive the transition from adenoma to adenocarcinoma. On the one hand, p53 mutations confer resistance to classical chemotherapy but, on the other hand, they open the door for immunotherapy, as p53-mutated tumors are rich in neoantigens. Aberrant function of the TP53 gene product, p53, also affects stromal and non-stromal cells in the tumor microenvironment. Cancer-associated fibroblasts together with other immunosuppressive cells become valuable assets for the tumor by p53-mediated tumor signaling. In this review, we address the manifold implications of p53 mutations in CRC regarding therapy, treatment response and personalized medicine.

Distinct functions of transforming growth factor-\u03b2 signaling in c-MYC driven hepatocellular carcinoma initiation and progression

Dysregulation of transforming growth factor-beta (TGFβ) signaling has been implicated in liver carcinogenesis with both tumor promoting and inhibiting activities. Activation of the c-MYC protooncogene is another critical genetic event in hepatocellular carcinoma (HCC). However, the precise functional crosstalk between c-MYC and TGFβ signaling pathways remains unclear. In the present investigation, we investigated the expression of TGFβ signaling in c-MYC amplified human HCC samples as well as the mechanisms whereby TGFβ modulates c-Myc driven hepatocarcinogenesis during initiation and progression. We found that several TGFβ target genes are overexpressed in human HCCs with c-MYC amplification. In vivo, activation of TGFβ1 impaired c-Myc murine HCC initiation, whereas inhibition of TGFβ pathway accelerated this process. In contrast, overexpression of TGFβ1 enhanced c-Myc HCC progression by promoting tumor cell metastasis. Mechanistically, activation of TGFβ promoted tumor microenvironment reprogramming rather than inducing epithelial-to-mesenchymal transition during HCC progression. Moreover, we identified PMEPA1 as a potential TGFβ1 target. Altogether, our data underline the divergent roles of TGFβ signaling during c-MYC induced HCC initiation and progression.

CRISPR/dCas9-mediated transposition with specificity and efficiency of site-directed genomic insertions.

The ability and efficiency of targeted nucleases to perform sequence replacements or insertions into the genome are limited. This limited efficiency for sequence replacements or insertions can be explained by the dependency on DNA repair pathways, the possibility of cellular toxicity, and unwanted activation of proto-oncogenes. The piggyBac (PB) transposase uses a very efficient enzymatic mechanism to integrate DNA fragments into the genome in a random manner. In this study, we fused an RNA-guided catalytically inactive Cas9 (dCas9) to the PB transposase and used dual sgRNAs to localize this molecule to specific genomic targets. We designed and used a promoter/reporter complementation assay to register and recover cells harboring-specific integrations, where only by complementation upon correct genomic integration, the reporter can be activated. Using an RNA-guided piggyBac transposase and dual sgRNAs, we were able to achieve site-directed integrations in the human ROSA26 safe harbor region in 0.32% of cells. These findings show that the methodology used in this study can be used for targeting genomic regions. An application for this finding could be in cancer cells to insert sequences into specific target regions that are intended to be destroyed, or to place promoter cargos behind the tumor suppressor genes to activate them.

Methylation alterations and advance of treatment in lymphoma.

Lymphoma is a common and aggressive form of hematopoietic malignancies with diverse clinical and pathological features due to its heterogeneity. Although the current immunochemotherapeutic regimens improve clinical outcomes, many patients still display poor prognosis and frequent relapse. Epigenetic alterations contribute to the progression of lymphoma. DNA methylation and histone methylation are the most common epigenetic alterations and regulate the gene expression involved in lymphoma pathogenesis, including silencing of tumor suppressor genes or activation of proto-oncogenes. Dysregulation or mutation of genes related to DNA methylation, including DNMTs, TET2, IDH2, and genes related to histone methylation, including EZH2, KMT2D has been observed. Most of these alterations are associated with inferior outcomes of patients with diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), peripheral T-cell lymphoma (PTCL), and other subtypes of lymphoma. To overcome the pathogenetic consequence induced by aberrant DNA methylation and histone methylation, novel targeted drugs including azacitidine and decitabine have been gradually applied in practice to enhance the efficacy of current therapy and improve the prognosis of lymphoma patients. Investigating and targeting epigenetic mechanisms in lymphoma could be a key point of future research. Therefore, we mainly summarize the methylation alterations in lymphoma and their respective targeted therapies in this review.

Molecular Mechanisms of Hepatoblastoma.

Hepatoblastoma (HB) is the predominant primary liver tumor in children. While the prognosis is favorable when the tumor can be resected, the outcome is dismal for patients with progressed HB. Therefore, a better understanding of the molecular mechanisms responsible for HB is imperative for early detection and effective treatment. Sequencing analysis of human HB specimens unraveled the pivotal role of Wnt/β-catenin pathway activation in this disease. Nonetheless, β-catenin activation alone does not suffice to induce HB, implying the need for additional alterations. Perturbations of several pathways, including Hippo, Hedgehog, NRF2/KEAP1, HGF/c-Met, NK-1R/SP, and PI3K/AKT/mTOR cascades and aberrant activation of c-MYC, n-MYC, and EZH2 proto-oncogenes, have been identified in HB, although their role requires additional investigation. Here, we summarize the current knowledge on HB molecular pathogenesis, the relevance of the preclinical findings for the human disease, and the innovative therapeutic strategies that could be beneficial for the treatment of HB patients.

STAT3 and its targeting inhibitors in osteosarcoma.

Signal transducer and activator of transcription 3 (STAT3) is one of seven STAT family members involved with the regulation of cellular growth, differentiation and survival. STAT proteins are conserved among eukaryotes and are important for biological functions of embryogenesis, immunity, haematopoiesis and cell migration. STAT3 is widely expressed and located in the cytoplasm in an inactive form. STAT3 is rapidly and transiently activated by tyrosine phosphorylation by a range of signalling pathways, including cytokines from the IL‐6 family and growth factors, such as EGF and PDGF. STAT3 activation and subsequent dimer formation initiates nuclear translocation of STAT3 for the regulation of target gene transcription. Four STAT3 isoforms have been identified, which have distinct biological functions. STAT3 is considered a proto‐oncogene and constitutive activation of STAT3 is implicated in the development of various cancers, including multiple myeloma, leukaemia and lymphomas. In this review, we focus on recent progress on STAT3 and osteosarcoma (OS). Notably, STAT3 is overexpressed and associated with the poor prognosis of OS. Constitutive activation of STAT3 in OS appears to upregulate the expression of target oncogenes, leading to OS cell transformation, proliferation, tumour formation, invasion, metastasis, immune evasion and drug resistance. Taken together, STAT3 is a target for cancer therapy, and STAT3 inhibitors represent potential therapeutic candidates for the treatment of OS.

Micro-RNA networks in T-cell prolymphocytic leukemia reflect T-cell activation and shape DNA damage response and survival pathways.

T-cell prolymphocytic leukemia (T-PLL) is a poor-prognostic mature T-cell malignancy. It typically presents with exponentially rising lymphocyte counts, splenomegaly, and bone marrow infiltration. Effective treatment options are scarce and a better understanding of TPLL’s pathogenesis is desirable. Activation of the TCL1 proto-oncogene and loss-of-function perturbations of the tumor suppressor ATM are TPLL’s genomic hallmarks. The leukemic cell reveals a phenotype of active T-cell receptor (TCR) signaling and aberrant DNA damage responses. Regulatory networks based on the profile of microRNA (miR) have not been described for T-PLL. In a combined approach of small-RNA and transcriptome sequencing in 46 clinically and moleculary well-characterized T-PLL, we identified a global T-PLL-specific miR expression profile that involves 34 significantly deregulated miR species. This pattern strikingly resembled miR-ome signatures of TCR-activated T cells. By integrating these T-PLL miR profiles with transcriptome data, we uncovered regulatory networks associated with cell survival signaling and DNA damage response pathways. Despite a miR-ome that discerned leukemic from normal T cells, there were also robust subsets of T-PLL defined by a small set of specific miR. Most prominently, miR-141 and the miR- 200c-cluster separated cases into two major subgroups. Furthermore, increased expression of miR-223-3p as well as reduced expression of miR-21 and the miR-29 cluster were associated with more activated Tcell phenotypes and more aggressive disease presentations. Based on the implicated pathobiological role of these miR deregulations, targeting strategies around their effectors appear worth pursuing. We also established a combinatorial miR-based overall survival score for T-PLL (miROS-T-PLL), that might improve current clinical stratifications.