Uncontrolled Cellular Proliferation Research Articles

Detection of Lung Cancer diseases Based on CT SCAN Images using Deep Learning

Abstract: Increased health risks originate from changes in the natural surroundings, climate, and way of life. India is the leading nation in respiratory illness mortality. They were always the second-leading mortality cause globally in 2017 behind heart disease, claiming the lives of 1 million (958,000) Indians. Severe effects of lung cancer, especially death, can be avoided by timely detection and treatment. The better diagnostic technique at the present is a chest X-ray, which is necessary for clinical therapy. By using Deep Learning to detect lung diseases from chest X-rays, a person with lung disease may be able to save their own life. Lung tumor is yet another name for lung cancer. It is a malignant tumor illness that causes uncontrolled cellular proliferation in the lung tissue. The much more common sources of lung cancer by just using tobacco products and smoking. One of the most common causes of death in the entire world is lung cancer. The ResNet-50 (Residual Network) pre-trained Deep Learning model used for image classification of the Convolutional Neural Network is a very successful method. Neural network (also known as Convolution or CNN). ResNet-50, which now has 50 layers, has been trained using just a million images from the ImageNet database in 1000 various categories. The ability to forecast outcomes quickly and accurately tends to make this possible. This paper presents a practical strategy for just using deep learning to recognize lung problems. Its primary objective is to develop a method that will assist radiologists to identify lung problems. This will be especially beneficial in remote locations in which radiologists are in short supply. To evaluate the efficacy and correctness of various models for trying to identify lung cancer from chest x-ray image data, the RESNET 50 model is used.

Precision oncology with selective RET inhibitor selpercatinib in RET-rearranged cancers.

Rearranged during transfection (RET) is a protooncogene that encodes for receptor tyrosine kinase with downstream effects on multiple cellular pathways. Activating RET alterations can occur and lead to uncontrolled cellular proliferation as a hallmark of cancer development. Oncogenic RET fusions are present in nearly 2% of patients with non-small cell lung cancer (NSCLC), 10-20% of patients with thyroid cancer, and <1% across the pan-cancer spectrum. In addition, RET mutations are drivers in 60% of sporadic medullary thyroid cancers and 99% of hereditary thyroid cancers. The discovery, rapid clinical translation, and trials leading to FDA approvals of selective RET inhibitors, selpercatinib and pralsetinib, have revolutionized the field of RET precision therapy. In this article, we review the current status on the use of the selective RET inhibitor, selpercatinib, in RET fusion-positive tumors: NSCLC, thyroid cancers, and the more recent tissue-agnostic activity leading to FDA approval.

BRAF/MEK inhibition in NSCLC: mechanisms of resistance and how to overcome it.

Targeted therapy for oncogenic genetic alterations has changed the treatment paradigm of advanced non-small cell lung cancer (NSCLC). Mutations in the BRAF gene are detected in approximately 4% of patients and result in hyper-activation of the MAPK pathway, leading to uncontrolled cellular proliferation. Inhibition of BRAF and its downstream effector MEK constitutes a therapeutic strategy for a subset of patients with NSCLC and is associated with clinical benefit. Unfortunately, the majority of patients will develop disease progression within 1year. Preclinical and clinical evidence suggests that resistance mechanisms involve the restoration of MAPK signaling which becomes inhibition-independent due to upstream or downstream alterations, and the activation of bypass pathways, such as the PI3/AKT/mTOR pathway. Future research should be directed to deciphering the mechanisms of cancer cells' oncogenic dependence, understanding the tissue-specific mechanisms of BRAF-mutant tumors, and optimizing treatment strategies after progression on BRAF and MEK inhibition.

Performing in situ hybridization and quantitative PCR to assess eya2 localization and expression in zebrafish

Rhabdomyosarcoma (RMS) is the most common soft‐tissue sarcoma found in children (Skapek, 2019) and is classified as a skeletal muscle cancer (Hsu, 2021). A common transcription factor that increases cell proliferation (how cells grow and divide) in RMS is called SIX1 (Hsu, 2021). The SIX1 protein has coregulators, such as EYA2, that regulate the increase or decrease of SIX1’s function (Sousounis, 2020). Because the coregulator EYA2 can play a critical role in the increase of SIX1's function (Blevins, 2015), we hypothesize that an increase in EYA2 will lead to uncontrolled cellular proliferation. This uncontrolled cellular proliferation could lead to the development of tumors, specifically RMS. In order to understand EYA2’s role in tumor growth and cancer, it is important to study the coregulator under normal homeostatic conditions. Specifically, understanding how muscle tissue normally utilizes eya2 during embryonic development, when cells are proliferating, can provide insight into eya2’s potential roles in muscle tumor development. Zebrafish are ideal research specimens for this research because of their easy to visualize muscular system. A technique used to visualize mRNA expression and localization in zebrafish tissues, is in situ hybridization (ISH) (Chu, 2019). Our first objective was to confirm that we can perform ISH in our laboratory. We used ISH to observe that Eya2 mRNA was generally located in the head and upper torso regions of the zebrafish at both 24 hours post fertilization (hpf) and 48 hpf, which supports previously published results (Thisse, 2004; Thisse, 2004). Additionally, we observed an increase in eya2 mRNA levels by ISH from 24 hpf to 48 hpf. We utilized quantitative PCR (qPCR) (Heid, 1996) to quantify these changes in eya2 mRNA levels. Our qPCR results indicate there was an increase in the coregulator eya2 mRNA expression from 24 hpf to 48 hpf. Overall, both research techniques, ISH and qPCR, demonstrated an increase in eya2 transcript levels from 24 hpf to 48 hpf, suggesting that this time frame would be appropriate for future studies of eya2 function in zebrafish development.

Small-Molecule RAS Inhibitors as Anticancer Agents: Discovery, Development, and Mechanistic Studies.

Mutations of RAS oncogenes are responsible for about 30% of all human cancer types, including pancreatic, lung, and colorectal cancers. While KRAS1 is a pseudogene, mutation of KRAS2 (commonly known as KRAS oncogene) is directly or indirectly associated with human cancers. Among the RAS family, KRAS is the most abundant oncogene related to uncontrolled cellular proliferation to generate solid tumors in many types of cancer such as pancreatic carcinoma (over 80%), colon carcinoma (40–50%), lung carcinoma (30–50%), and other types of cancer. Once described as ‘undruggable’, RAS proteins have become ‘druggable’, at least to a certain extent, due to the continuous efforts made during the past four decades. In this account, we discuss the chemistry and biology (wherever available) of the small-molecule inhibitors (synthetic, semi-synthetic, and natural) of KRAS proteins that were published in the past decades. Commercial drugs, as well as investigational molecules from preliminary stages to clinical trials, are categorized and discussed in this study. In summary, this study presents an in-depth discussion of RAS proteins, classifies the RAS superfamily, and describes the molecular mechanism of small-molecule RAS inhibitors

Beyond Good and Evil: Molecular Mechanisms of Type I and III IFN Functions.

IFNs are comprised of three families of cytokines that confer protection against pathogen infection and uncontrolled cellular proliferation. The broad role IFNs play in innate and adaptive immune regulation has placed them under heavy scrutiny to position them as "friend" or "foe" across pathologies. Genetic lesions in genes involving IFN synthesis and signaling underscore the disparate outcomes of aberrant IFN signaling. Abrogation of the response leads to susceptibility to microbial infections whereas unabated IFN induction underlies a variety of inflammatory diseases and tumor immune evasion. Type I and III IFNs have overlapping roles in antiviral protection, yet the mechanisms by which they are induced and promote the expression of IFN-stimulated genes and inflammation can distinguish their biological functions. In this review, we examine the molecular factors that shape the shared and distinct roles of type I and III IFNs in immunity.

Computational modeling of potential milciclib derivatives inhibitor-CDK2 binding through global docking and accelerated molecular dynamics simulations

Hepatocellular carcinoma (HCC) is the most common malignant condition of the liver that occurs as a result of uncontrolled cellular proliferation after a series of disruptions at cell cycle regulatory checkpoints in the normal cell. Due to the lack of appropriate therapeutics or remedial treatment methods, new treatment strategies against HCC need to be developed. Cyclin dependent kinases (CDKs) are required to control the cell cycle and apoptosis, but their overexpressionis critical in the progression of cancer and is often expressed in HCC. Thus, CDKs are considered a promising class of target-defined therapy for HCC. Milciclib is a potential candidate for HCC which exhibits inhibitory activity against CDK2 leading to cell cycle arrest and apoptosis of tumor cells. Herein, we have halogenated the parent drug milciclib to improve its efficacy against CDK2. The primary structure of milciclib (D) was modified with F, Cl and,CF3 groups. The frontier molecular orbital features, binding affinity, non-bonded interaction and the pharmacokinetic parameters were analyzed for milciclib and its derivatives. We also performed molecular docking and extended molecular dynamics simulation studies to study the binding interactions and binding affinity more closely. Our computational investigation showed the derivatives D-F and D-CF3 have significant chemical reactivity, the best binding affinity, nonbonding interactions, and improved pharmacokinetic properties compared to the parent drug milciclib. Molecular dynamics analysis and MM-PBSA calculations indicated that D-Cl had a slightly more stable conformation and higher binding affinity compared to D-CF3.

Nuclear Receptor Coactivator NCOA3 Regulates UV Radiation-Induced DNA Damage and Melanoma Susceptibility.

Melanoma occurs as a consequence of inherited susceptibility to the disease and exposure to UV radiation (UVR) and is characterized by uncontrolled cellular proliferation and a high mutational load. The precise mechanisms by which UVR contributes to the development of melanoma remain poorly understood. Here we show that activation of nuclear receptor coactivator 3 (NCOA3) promotes melanomagenesis through regulation of UVR sensitivity, cell-cycle progression, and circumvention of the DNA damage response (DDR). Downregulation of NCOA3 expression, either by genetic silencing or small-molecule inhibition, significantly suppressed melanoma proliferation in melanoma cell lines and patient-derived xenografts. NCOA3 silencing suppressed expression of xeroderma pigmentosum C and increased melanoma cell sensitivity to UVR. Suppression of NCOA3 expression led to activation of DDR effectors and reduced expression of cyclin B1, resulting in G2-M arrest and mitotic catastrophe. A SNP in NCOA3 (T960T) reduced NCOA3 protein expression and was associated with decreased melanoma risk, given a significantly lower prevalence in a familial melanoma cohort than in a control cohort without cancer. Overexpression of wild-type NCOA3 promoted melanocyte survival following UVR and was accompanied by increased levels of UVR-induced DNA damage, both of which were attenuated by overexpression of NCOA3 (T960T). These results describe NCOA3-regulated pathways by which melanoma can develop, with germline NCOA3 polymorphisms enabling enhanced melanocyte survival in the setting of UVR exposure, despite an increased mutational burden. They also identify NCOA3 as a novel therapeutic target for melanoma. SIGNIFICANCE: This study explores NCOA3 as a regulator of the DDR and a therapeutic target in melanoma, where activation of NCOA3 contributes to melanoma development following exposure to ultraviolet light.

Expression of p53 and Ki-67 proteins in patients with increasing severity and duration of pterygium.

Purpose:Pterygium is a triangular fibrovascular subepithelial ingrowth of degenerative bulbar conjunctival tissue over the cornea. It is now considered to be a result of uncontrolled cellular proliferation as overexpression of p53 protein and Ki-67 nuclear protein was found in the epithelium. This study was done to find the expression of p53 and Ki-67 with the severity and duration of the pterygium to explain the etiopathogenesis.Methods:Data were analyzed from 43 Indian participants of all age groups. All patients were divided according to the severity of pterygium (mild, moderate, and severe groups) and according to the duration of pterygium (<4 years and >4 years). The samples were studied by immunohistochemistry by using antibodies against p53 and Ki-67 proteins considering >5% expression as significant.Results:Of 43 cases, p53 and Ki-67 expression were positive in 33 cases. In mild, moderate, and severe cases p53 positivity was 33.3%, 78.4%, 100%, respectively. P53 expression increased with duration, 79.3% positive in <4 years, and 92.9% positive in >4 years. With increasing severity of pterygium, mild, moderate, and severe cases, Ki-67 positivity was 66.7%, 78.37%, 66.7%, respectively. Ki-67 expression with duration, 79.3% positive in <4 years, and 85.7% positive in >4 years of the duration of pterygium with no statistical significance.Conclusion:Our study revealed that with increasing duration and severity of pterygium, p53 expression was observed to be increasing. Ki-67 expression increased with the duration of pterygium but not with the severity.

Multi-institutional noninvasive in vivo characterization of IDH, 1p/19q, and EGFRvIII in glioma using neuro-Cancer Imaging Phenomics Toolkit (neuro-CaPTk).

BackgroundGliomas represent a biologically heterogeneous group of primary brain tumors with uncontrolled cellular proliferation and diffuse infiltration that renders them almost incurable, thereby leading to a grim prognosis. Recent comprehensive genomic profiling has greatly elucidated the molecular hallmarks of gliomas, including the mutations in isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2), loss of chromosomes 1p and 19q (1p/19q), and epidermal growth factor receptor variant III (EGFRvIII). Detection of these molecular alterations is based on ex vivo analysis of surgically resected tissue specimen that sometimes is not adequate for testing and/or does not capture the spatial tumor heterogeneity of the neoplasm.MethodsWe developed a method for noninvasive detection of radiogenomic markers of IDH both in lower-grade gliomas (WHO grade II and III tumors) and glioblastoma (WHO grade IV), 1p/19q in IDH-mutant lower-grade gliomas, and EGFRvIII in glioblastoma. Preoperative MRIs of 473 glioma patients from 3 of the studies participating in the ReSPOND consortium (collection I: Hospital of the University of Pennsylvania [HUP: n = 248], collection II: The Cancer Imaging Archive [TCIA; n = 192], and collection III: Ohio Brain Tumor Study [OBTS, n = 33]) were collected. Neuro-Cancer Imaging Phenomics Toolkit (neuro-CaPTk), a modular platform available for cancer imaging analytics and machine learning, was leveraged to extract histogram, shape, anatomical, and texture features from delineated tumor subregions and to integrate these features using support vector machine to generate models predictive of IDH, 1p/19q, and EGFRvIII. The models were validated using 3 configurations: (1) 70–30% training–testing splits or 10-fold cross-validation within individual collections, (2) 70–30% training–testing splits within merged collections, and (3) training on one collection and testing on another.ResultsThese models achieved a classification accuracy of 86.74% (HUP), 85.45% (TCIA), and 75.15% (TCIA) in identifying EGFRvIII, IDH, and 1p/19q, respectively, in configuration I. The model, when applied on combined data in configuration II, yielded a classification success rate of 82.50% in predicting IDH mutation (HUP + TCIA + OBTS). The model when trained on TCIA dataset yielded classification accuracy of 84.88% in predicting IDH in HUP dataset.ConclusionsUsing machine learning algorithms, high accuracy was achieved in the prediction of IDH, 1p/19q, and EGFRvIII mutation. Neuro-CaPTk encompasses all the pipelines required to replicate these analyses in multi-institutional settings and could also be used for other radio(geno)mic analyses.

Cyclin D1 in Cancer: A Molecular Connection for Cell Cycle Control, Adhesion and Invasion in Tumor and Stroma.

Cyclin D1, an important regulator of cell cycle, carries out a central role in the pathogenesis of cancer determining uncontrolled cellular proliferation. In normal cells, Cyclin D1 expression levels are strictly regulated, conversely, in cancer, its activity is intensified in various manners. Different studies demonstrate that CCDN1 gene is amplified in several tumor types considering it as a negative prognostic marker of this pathology. Cyclin D1 is known for its role in the nucleus, but recent clinical studies associate the amount located in the cytoplasmic membrane with tumor invasion and metastasis. Cyclin D1 has also other functions: it governs the expression of specific miRNAs and it plays a crucial role in the tumor-stroma interactions potentiating most of the cancer hallmarks. In the present review, we will summarize the current scientific evidences that highlight the involvement of Cyclin D1 in the pathogenesis of different types of cancer, best of all in breast cancer. We will also focus on recent insights regarding the Cyclin D1 as molecular bridge between cell cycle control, adhesion, invasion, and tumor/stroma/immune-system interplay in cancer.

Engineered Nanoparticles for Cancer Vaccination and Immunotherapy.

The immune system has evolved over time to protect the host from foreign microorganisms. Activation of the immune system is predicated on a distinction between self and nonself. Unfortunately, cancer is characterized by genetic alterations in the host's cells, leading to uncontrolled cellular proliferation and evasion of immune surveillance. Cancer immunotherapy aims to educate the host's immune system to not only recognize but also attack and kill mutated cancer cells. While immune checkpoint blockers have been proven to be effective against multiple types of advanced cancer, the overall patient response rate still remains below 30%. Therefore, there is an urgent need to improve current cancer immunotherapies. In this Account, we present an overview of our recent progress on nanoparticle-based strategies for improving cancer vaccines and immunotherapies. We also present other complementary strategies to give a well-rounded snapshot of the field of combination cancer immunotherapy. The versatility and tunability of nanoparticles make them promising platforms for addressing individual challenges posed by various cancers. For example, nanoparticles can deliver cargo materials to specific cells, such as vaccines delivered to antigen-presenting cells for strong immune activation. Nanoparticles also allow for stimuli-responsive delivery of various therapeutics to cancer cells, thus forming the basis for combination cancer immunotherapy. Here, we focus on nanoparticle platforms engineered to deliver tumor antigens, whole tumor cells, and chemotherapeutic or phototherapeutic agents in a manner to effectively and safely trigger the host's immune system against tumor cells. For each work, we discuss the nanoparticle platform developed, synthesis chemistry, and in vivo applications. Nanovaccines offer a unique platform for codelivery of personalized tumor neoantigens and adjuvants and elicitation of robust immune responses against aggressive tumors. Nanovaccines either delivering whole tumor cell lysate or formed from tumor cell lysate may increase the repertoire of tumor antigens as immune targets while exploiting immunogenic cell death to prime antitumor immune responses. We also discuss how antigen- and whole tumor cell-based approaches may open the door for personalized cancer vaccination and immunotherapy. On the other hand, chemotherapy, phototherapy, and radiotherapy are more standardized cancer therapies, and nanoparticle-based approaches may promote their ability to initiate T cell activation against tumor cells and improve antitumor efficacy with minimal toxicity. Finally, building on the recent progress in nanoparticle-based cancer immunotherapy, the field should set the ultimate goal to be clinical translation and clinical efficacy. We will discuss regulatory, analytical, and manufacturing hurdles that should be addressed to expedite the clinical translation of nanomedicine-based cancer immunotherapy.

Neurofibromatosis Type 1: Ocular Electrophysiological and Perimetric Anomalies.

IntroductionNeurofibromatosis type 1 (NF1) is a multisystemic disease caused by the mutation of Nf1 gene located on chromosome 17q11.2. The mutation determines the loss of function of the protein neurofibromin with consequent uncontrolled cellular proliferation. Patients are characterized by a wide range of dermatological, neurological, and ophthalmological symptoms.PurposeThe aim of the study was to evaluate, through pattern visual evoked potentials (p-VEPs) and frequency doubling technology (FDT) Matrix perimetry, the objective and psychophysical functionality of the optic pathways in a group of NF1 patient.MethodsThe study group consisted of 26 patients affected by NF1 and 17 healthy controls. Each patient underwent a complete ophthalmological examination, p-VEPs with the evaluation of amplitude and latency of the P100 wave, and FDT perimetry, with the evaluation of central sensitivity (CS), mean deviation (MD), pattern standard deviation (PSD) and glaucoma hemifield test (GHT).ResultsNF1 patients showed a statistically significant alteration in the transmission of visual impulse. P-VEPs results highlighted a reduced amplitude and an increased latency of the P100 wave, suggesting an involvement of the visual pathway. Visual field analysis showed a significant reduction in all the observed parameters as well (CS, MD, PSD, and GHT).ConclusionThe present study showed, in NF1 patients, a qualitative and quantitative alteration in the conduction of stimuli through the visual pathways. The observed alterations are present, although, only at a subclinical level. None of the patients included in the study showed any manifest visual deficit nor had any concomitant pathology that might have affected the outcome of the study. In conclusion, electrophysiological exams and computer perimetry may take part, alongside a wider array of exams, in the differential diagnosis and later monitoring of NF1.

An Intimate Alliance of DNA-Damage Response Network with Cell-Cycle Checkpoints Amid Events of Uncontrolled Cellular Proliferation: A Mini- Review.

There is close interdependence between cell survival, cell senescence, events of the cell cycle, apoptosis, malignancy development, and tumor responses to cancer treatment. Intensive studies and elaborate researches have been conducted on the functional aspects of oncogenes, tumor suppressor genes, apoptotic genes, and members guiding cell cycle regulation. These disquisitions have put forward the existence of a highly organized response pathway termed as a DNA-damage response network. The pathways detecting DNA damage and signaling are intensively linked to the events of cell-cycle arrest, cell proliferation, apoptosis, and cell senescence. DNA damage responses are complex systems that incorporate specific "sensor" and "transducer" proteins, for assessment of damage and signal transmission, respectively. These signals are thereafter relayed upon various "effector" proteins involved in different cellular pathways. It may include those governing cell-cycle checkpoints, participating in DNA repair, cell senescence, and apoptosis. This review discusses the role of the tumour suppressor gene, oncogenes, cell cycle checkpoint regulators during DNA damage response and regulation.

Abstract 3963: Physical confinement induces malignant transformation in mammary epithelial cells

Abstract The physical microenvironment of tumor cells plays an important role in cancer initiation and progression. Here, we present evidence that confinement - a new physical parameter that is apart from matrix stiffness - can also induce malignant transformation in mammary epithelial cells. We discovered that MCF10A cells, a benign mammary cell line that forms growth-arrested polarized acini in Matrigel, transforms into cancer-like cells within the same Matrigel material following confinement in alginate shell hydrogel microcapsules. The confined cells exhibited a range of tumor-like behaviors, including uncontrolled cellular proliferation and invasion. Additionally, 4-6 weeks after transplantation into the mammary fad pads of immunocompromised mice, the confined cells formed large palpable masses that exhibited histological features similar to that of carcinomas. Taken together, our findings suggest that physical confinement represents a previously unrecognized mechanism for malignancy induction in mammary epithelial cells and also provide a new, microcapsule-based, high throughput model system for testing new breast cancer therapeutics. Citation Format: Yen-Chun Lu. Physical confinement induces malignant transformation in mammary epithelial cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3963.

Ocular surface squamous neoplasia: Population demographics, pathogenesis and risk factors

Background: Ocular surface squamous neoplasia (OSSN) is a unifying term used to describe conjunctival intra-epithelial neoplasia, squamous cell carcinoma in situ and invasive squamous cell carcinoma.Aim: The aim of this article was to describe the demographics, clinical features, pathogenesis and risk factors of OSSN.Method: A literature search was conducted using the search criteria ‘ocular surface squamous neoplasia’, ‘diagnosis’, ‘epidemiology’, ‘pathogenesis’ and ‘risk factors’.Results: Ocular surface squamous neoplasia is the most common ocular tumour, with incidence rates ranging from 0.01 to 3.4 per 100 000 persons/year. There are two main patterns of disease presentation: older white males in temperate climates where human immunodeficiency virus (HIV) and human papilloma virus (HPV) are not associated; and a younger patient population in tropical climates where HIV and HPV are more prevalent. The pathogenesis primarily revolves around ultraviolet B exposure and HPV infection that cause genetic mutations and uncontrolled cellular proliferation, whilst HIV infection and vitamin A impair tumour surveillance mechanisms. Ocular surface squamous neoplasia is first suspected clinically before formal confirmation of the diagnosis. Morphologically, it can be divided into three groups: placoid, nodular and diffuse. Placoid lesions can further be sub-divided into gelatinous, leukoplakic and papilliform lesions. Nodular lesions have the poorest prognosis, with the highest risk of metastasis and recurrence.Conclusion: Ocular surface squamous neoplasia is a common ocular tumour associated with ultraviolet radiation, HPV and HIV infection. The pathogenesis revolves around acquired genetic mutations, unregulated cellular proliferation and impaired tumour surveillance mechanisms.

E2F5 promotes prostate cancer cell migration and invasion through regulation of TFPI2, MMP-2 and MMP-9.

Previously, our laboratory demonstrated that a deregulated E2F5/p38/SMAD3 axis was associated with uncontrolled cellular proliferation in prostate cancer (PCa). Here, we investigate the role of E2F5 in PCa in further details. RNAi-mediated E2F5 knockdown and pathway-focused gene expression profiling in PC3 cells identified TFPI2 as a downstream target of E2F5. Manipulation of E2F5 expression was also found to alter MMP-2 and MMP-9 levels as detected by Proteome Profiler array, western blot and reverse transcription coupled quantitative polymerase chain reaction Site-directed mutagenesis, dual-luciferase assays and chromatin immunoprecipitation with anti-E2F5-IgG coupled with qPCR confirmed recruitment of E2F5 on TFPI2, MMP-2 and MMP-9 promoters. RNAi-mediated knockdown of E2F5 expression in PC3 caused a significant alteration of cell migration while that of TFFI2 resulted in a modest change. Abrogation of E2F5 and TFPI2 expression was associated with significant changes in the gelatinolytic activity of active forms of MMP-2 and MMP-9. Moreover, E2F5, MMP-2 and MMP-9 levels were elevated in biopsies of PCa patients relative to that of benign hyperplasia, while TFPI2 expression was reduced. MMP-9 was coimmunoprecipitated with anti-TFPI2-IgG in PCa tissue samples suggesting a direct interaction between the proteins. Finally, artemisinin treatment in PC3 cells repressed E2F5 along with MMP-2/MMP-9 while triggering TFPI2 expression which alleviated PC3 aggressiveness possibly through inhibition of MMP activities. Together, our study reinstates an oncogenic role of E2F5 which operates as a dual-function transcription factor for its targets TFPI2, MMP-2 and MMP-9 and promotes cellular invasiveness. This study also indicates a therapeutic potential of artemisinin, a natural compound which acts by correcting dysfunctional E2F5/TFPI2/MMP axis in PCa.

Gene Networks with Transcriptional Bursting Recapitulate Rare Transient Coordinated High Expression States in Cancer.

Non-genetic transcriptional variability is a potential mechanism for therapy resistance in melanoma. Specifically, rare subpopulations of cells occupy a transient pre-resistant state characterized by coordinated high expression of several genes and survive therapy. How might these rare states arise and disappear within the population? It is unclear whether the canonical models of probabilistic transcriptional pulsing can explain this behavior, or if it requires special, hitherto unidentified mechanisms. We show that a minimal model of transcriptional bursting and gene interactions can give rise to rare coordinated high expression states. These states occur more frequently in networks with low connectivity and depend on three parameters. While entry into these states is initiated by a long transcriptional burst that also triggers entry of other genes, the exit occurs through independent inactivation of individual genes. Together, we demonstrate that established principles of gene regulation are sufficient to describe this behavior and argue for its more general existence. A record of this paper's transparent peer review process is included in the Supplemental Information.

Tempol Treatment Inhibits Inflammatory Process and Improves Morphological Aspects in Early and Late‐stage of Prostate Cancer Progression in the Ventral Lobe of TRAMP Model.

Tempol (4‐hydroxy‐TEMPO) is a SOD mimetic and member of the nitroxide compounds family. It is used as a co‐adjuvant or protective factor in cancer treatment due to its anti‐oxidant properties. However, in prostate cancer (PCa) Tempol effects are still poorly understood. Its known that PCa progression requires a sustained inflammation, which generates a broad spectrum of reactive nitrogen and oxygen species, leading to uncontrolled cellular proliferation. We aimed to evaluate the Tempol effects in different stages of PCa progression (a) in vivo using a transgenic adenocarcinoma of the mouse prostate model (TRAMP); (b) and in vitro using human PCa cell lines with different genetic background. For the early‐stage analysis, 8‐weeks‐old mice (TPL1 group) were treated orally with 50 mg/kg of Tempol during 4 weeks. For the late‐stage analysis, 16‐weeks‐old mice were treated with 50mg/Kg (TPL2 group) or 100mg/Kg (TPL3 group) of Tempol during the same period. All control groups received water as vehicle. The ventral lobe of the prostate was collected and submitted to the histopathological, immunohistochemical and Western blotting analysis. PC‐3 and LnCaP cells were exposed to different doses of Tempol (0.5, 1.0 and 2.0 mM) and cell viability assay was performed. In vivo, Tempol treatment promoted a great improvement in the prostatic epithelium in all the analyzed cancer stages, reducing significantly premalignant (high‐grade prostatic intraepithelial neoplasia) and malignant lesions (well differentiated prostatic adenocarcinoma). The PCNA immunostaining confirmed the cellular proliferation decrease between the control and treated groups. Tempol also reduced COX‐2 and NFkB, especially in the early‐stage, whereas iNOS increased in this period. In the late‐stage, both doses of Tempol reduced NFkB levels concomitant with IkB‐α increase, suggesting an inhibitory effect of Tempol on NFkB pathway in association with its anti‐oxidant role. In vitro, Tempol decreased cell viability of both cell lines, being the 1.0 mM dose the most efficient and nontoxic concentration. In conclusion, Tempol had a remarkable action in the prostate, delaying cancer progression in the ventral lobe of TRAMP model and inhibiting PC‐3 and LnCaP growth. This SOD mimetic presented great capability of inhibiting the inflammation process, and consequently the cell proliferation. Its action was effective in different stages of PCa progression, particularly in the early stage, indicating that Tempol treatment can be explored as a preventive approach. Finally, for the late PCa stage, Tempol may be considered as an interesting adjuvant therapy.Support or Funding InformationSão Paulo Research Foundation, FAPESP, process 2018/21647‐6 and FAEPEX/UNICAMP, process 2202/19

Oral Squamous Cell Carcinoma and the Cutting Edge of Nanotechnology

Introduction: Cancer is a group of diseases characterized by uncontrolled cellular growth or proliferation. Oral squamous cell carcinoma (OSCC) is a group of cancers mainly affecting the buccal cavity and associated area, with high rate of mortality and morbidity. Chronic consumption of alcohol, both smoking and smokeless tobacco, and betel quid are considered as the prominent determinants for the initiation and development of OSCC. The current treatment strategy of OSCC is associated with the common side effects of non-specific anticancer drug delivery. In addition, the treatment also deals with the recurrence of OSCC after completion of the therapy, which is responsible for its high mortality rate. Nanotechnology is an emerging discipline in drug delivery, disease diagnosis, and imaging. Nanoparticles are a group of colloidal carriers with approximately 1-1000 nm size range. Nanoparticles, due to large surface area to volume ratio, display different physicochemical characteristics favorable for their use in chemotherapy. The nanoparticles with options for modifications in theirsizes, surface charges, drug payload, and drug bioavailability can be used as a drug delivery system. The dynamic and flexible nature of nanoparticles can be employed to both active and passive targeting of cancer cells combating the usual side effects as well as other complexities associated with the conventional chemotherapy. Conclusions: The nanoparticles can be regarded as the future of drug delivery in cancer therapy.