Differentiation signals induce APOBEC3A expression via GRHL3 in squamous epithelia and squamous cell carcinoma.

Two APOBEC (apolipoprotein-B mRNA editing enzyme catalytic polypeptide-like) DNA cytosine deaminase enzymes (APOBEC3A and APOBEC3B) generate somatic mutations in cancer, driving tumour development and drug resistance. Here we used single cell RNA sequencing to study APOBEC3A and APOBEC3B expression in healthy and malignant mucosal epithelia, validating key observations with immunohistochemistry, spatial transcriptomics and functional experiments. Whereas APOBEC3B is expressed in keratinocytes entering mitosis, we show that APOBEC3A expression is confined largely to terminally differentiating cells and requires Grainyhead-like transcription factor 3 (GRHL3). Thus, in normal tissue, neither deaminase appears to be expressed at high levels during DNA replication, the cell cycle stage associated with APOBEC-mediated mutagenesis. In contrast, we show that in squamous cell carcinoma tissues, there is expansion of GRHL3 expression and activity to a subset of cells undergoing DNA replication and concomitant extension of APOBEC3A expression to proliferating cells. These findings indicate a mechanism for acquisition of APOBEC3A mutagenic activity in tumours.

Cardiovascular disease and chimeric antigen receptor cellular therapy.

Chimeric antigen receptor T-cell (CAR T) therapy is a revolutionary personalized therapy that has significantly impacted the treatment of patients with hematologic malignancies refractory to other therapies. Cytokine release syndrome (CRS) is a major side effect of CAR T therapy that can occur in 70-90% of patients, with roughly 40% of patients at grade 2 or higher. CRS can cause an intense inflammatory state leading to cardiovascular complications, including troponin elevation, arrhythmias, hemodynamic instability, and depressed left ventricular systolic function. There are currently no standardized guidelines for the management of cardiovascular complications due to CAR T therapy, but systematic practice patterns are emerging. In this review, we contextualize the history and indications of CAR T cell therapy, side effects related to this treatment, strategies to optimize the cardiovascular health prior to CAR T and the management of cardiovascular complications related to CRS. We analyze the existing data and discuss potential future approaches.

Gender Differences in Diagnosis, Prevention, and Treatment of Cardiotoxicity in Cardio-Oncology.

Gender differences exist throughout the medical field and significant progress has been made in understanding the effects of gender in many aspects of healthcare. The field of cardio-oncology is diverse and dynamic with new oncologic and cardiovascular therapies approved each year; however, there is limited knowledge regarding the effects of gender within cardio-oncology, particularly the impact of gender on cardiotoxicities. The relationship between gender and cardio-oncology is unique in that gender likely affects not only the biological underpinnings of cancer susceptibility, but also the response to both oncologic and cardiovascular therapies. Furthermore, gender has significant socioeconomic and psychosocial implications which may impact cancer and cardiovascular risk factor profiles, cancer susceptibility, and the delivery of healthcare. In this review, we summarize the effects of gender on susceptibility of cancer, response to cardiovascular and cancer therapies, delivery of healthcare, and highlight the need for further gender specific studies regarding the cardiovascular effects of current and future oncological treatments.

Cancer cell states recur across tumor types and form specific interactions with the tumor microenvironment.

Transcriptional heterogeneity among malignant cells of a tumor has been studied in individual cancer types and shown to be organized into cancer cell states; however, it remains unclear to what extent these states span tumor types, constituting general features of cancer. Here, we perform a pan-cancer single-cell RNA-sequencing analysis across 15 cancer types and identify a catalog of gene modules whose expression defines recurrent cancer cell states including 'stress', 'interferon response', 'epithelial-mesenchymal transition', 'metal response', 'basal' and 'ciliated'. Spatial transcriptomic analysis linked the interferon response in cancer cells to T cells and macrophages in the tumor microenvironment. Using mouse models, we further found that induction of the interferon response module varies by tumor location and is diminished upon elimination of lymphocytes. Our work provides a framework for studying how cancer cell states interact with the tumor microenvironment to form organized systems capable of immune evasion, drug resistance and metastasis.

Cancer cell states and emergent properties of the dynamic tumor system.

Phenotypic heterogeneity within malignant cells of a tumor is emerging as a key property of tumorigenesis. Recent work using single-cell transcriptomics has led to the identification of distinct cancer cell states across a range of cancer types, but their functional relevance and the advantage that they provide to the tumor as a system remain elusive. We present here a definition of cancer cell states in terms of coherently and differentially expressed gene modules and review the origins, dynamics, and impact of states on the tumor system as a whole. The spectrum of cell states taken on by a malignant population may depend on cellular lineage, epigenetic history, genetic mutations, or environmental cues, which has implications for the relative stability or plasticity of individual states. Finally, evidence has emerged that malignant cells in different states may cooperate or compete within a tumor niche, thereby providing an evolutionary advantage to the tumor through increased immune evasion, drug resistance, or invasiveness. Uncovering the mechanisms that govern the origin and dynamics of cancer cell states in tumorigenesis may shed light on how heterogeneity contributes to tumor fitness and highlight vulnerabilities that can be exploited for therapy.

Cooperation between melanoma cell states promotes metastasis through heterotypic cluster formation.

Melanomas can have multiple coexisting cell states, including proliferative (PRO) versus invasive (INV) subpopulations that represent a "go or grow" trade-off; however, how these populations interact is poorly understood. Using a combination of zebrafish modeling and analysis of patient samples, we show that INV and PRO cells form spatially structured heterotypic clusters and cooperate in the seeding of metastasis, maintaining cell state heterogeneity. INV cells adhere tightly to each other and form clusters with a rim of PRO cells. Intravital imaging demonstrated cooperation in which INV cells facilitate dissemination of less metastatic PRO cells. We identified the TFAP2 neural crest transcription factor as a master regulator of clustering and PRO/INV states. Isolation of clusters from patients with metastatic melanoma revealed a subset with heterotypic PRO-INV clusters. Our data suggest a framework for the co-existence of these two divergent cell populations, in which heterotypic clusters promote metastasis via cell-cell cooperation.

Exploring tissue architecture using spatial transcriptomics.

Deciphering the principles and mechanisms by which gene activity orchestrates complex cellular arrangements in multicellular organisms has far-reaching implications for research in the life sciences. Recent technological advances in next-generation sequencing- and imaging-based approaches have established the power of spatial transcriptomics to measure expression levels of all or most genes systematically throughout tissue space, and have been adopted to generate biological insights in neuroscience, development and plant biology as well as to investigate a range of disease contexts, including cancer. Similar to datasets made possible by genomic sequencing and population health surveys, the large-scale atlases generated by this technology lend themselves to exploratory data analysis for hypothesis generation. Here we review spatial transcriptomic technologies and describe the repertoire of operations available for paths of analysis of the resulting data. Spatial transcriptomics can also be deployed for hypothesis testing using experimental designs that compare time points or conditions-including genetic or environmental perturbations. Finally, spatial transcriptomic data are naturally amenable to integration with other data modalities, providing an expandable framework for insight into tissue organization.

Effect of yoga therapy on physical and psychological quality of life of perimenopausal women in selected coastal areas of Karnataka, India.

BACKGROUND: Perimenopausal period is characterized by a continuous decline in ovarian function due to which women are vulnerable to various physical and psychological symptoms affecting their quality of life. Currently these symptoms are managed by hormone replacement therapy. However, hormonal therapy can cause complications including malignancy which has resulted in search for various alternative therapies to improve the quality of life (QOL). Yoga is one such alternative therapy shown to enhance the QOL at all stages of human life associated with the chronic illness. There are very few scientific studies regarding the effect of yoga on perimenopause and in this study we investigated the effects of yoga therapy on physical and psychological symptoms using the standardized questionnaire. OBJECTIVE: To study the effect of yoga therapy on physical, psychological, vasomotor and sexual symptoms of perimenopause. MATERIALS AND METHODS: It is a prospective non-randomized control study of 216 perimenopausal women with 12 weeks of intervention. The subjects were divided in two groups with either yoga therapy [n = 111] or exercise [n = 105] as the interventional tool. The symptoms control and QOL before and after intervention in both the groups were assessed by using the menopausal QOL questionnaire. RESULTS: The perimenopausal symptoms in all the four domains were improved by yoga therapy, thus significantly improving the overall QOL compared to the control group. CONCLUSION: This study clearly demonstrates the effectiveness of yoga therapy in managing the distressing perimenopausal symptoms. It is easy, safe, non-expensive alternative therapy helping the well-being of perimenopausal women and must be encouraged in the regular management of perimenopausal symptoms.

A population of selected renal cells augments renal function and extends survival in the ZSF1 model of progressive diabetic nephropathy.

New treatment paradigms that slow or reverse progression of chronic kidney disease (CKD) are needed to relieve significant patient and healthcare burdens. We have shown that a population of selected renal cells (SRCs) stabilized disease progression in a mass reduction model of CKD. Here, we further define the cellular composition of SRCs and apply this novel therapeutic approach to the ZSF1 rat, a model of severe progressive nephropathy secondary to diabetes, obesity, dyslipidemia, and hypertension. Injection of syngeneic SRCs into the ZSF1 renal cortex elicited a regenerative response that significantly improved survival and stabilized disease progression to renal structure and function beyond 1 year posttreatment. Functional improvements included normalization of multiple nephron structures and functions including glomerular filtration, tubular protein handling, electrolyte balance, and the ability to concentrate urine. Improvements to blood pressure, including reduced levels of circulating renin, were also observed. These functional improvements following SRC treatment were accompanied by significant reductions in glomerular sclerosis, tubular degeneration, and interstitial inflammation and fibrosis. Collectively, these data support the utility of a novel renal cell-based approach for slowing renal disease progression associated with diabetic nephropathy in the setting of metabolic syndrome, one of the most common causes of end-stage renal disease.

Hedgehog-Gli pathway activation during kidney fibrosis.

The Hedgehog (Hh) signaling pathway regulates tissue patterning during development, including patterning and growth of limbs and face, but whether Hh signaling plays a role in adult kidney remains undefined. In this study, using a panel of hedgehog-reporter mice, we show that the two Hh ligands (Indian hedgehog and sonic hedgehog ligands) are expressed in tubular epithelial cells. We report that the Hh effectors (Gli1 and Gli2) are expressed exclusively in adjacent platelet-derived growth factor receptor-ß-positive interstitial pericytes and perivascular fibroblasts, suggesting a paracrine signaling loop. In two models of renal fibrosis, Indian Hh ligand was upregulated with a dramatic activation of downstream Gli effector expression. Hh-responsive Gli1-positive interstitial cells underwent 11-fold proliferative expansion during fibrosis, and both Gli1- and Gli2-positive cells differentiated into α-smooth muscle actin-positive myofibroblasts. In the pericyte-like cell line 10T1/2, hedgehog ligand triggered cell proliferation, suggesting a possible role for this pathway in the regulation of cell cycle progression of myofibroblast progenitors during the development of renal fibrosis. The hedgehog antagonist IPI-926 abolished Gli1 induction in vivo but did not decrease kidney fibrosis. However, the transcriptional induction of Gli2 was unaffected by IPI-926, suggesting the existence of smoothened-independent Gli activation in this model. This study is the first detailed description of paracrine hedgehog signaling in adult kidney, which indicates a possible role for hedgehog-Gli signaling in fibrotic chronic kidney disease.

Spectrophotometric analysis of the expression of secreted aspartyl proteinases from Candida in leukoplakia and oral squamous cell carcinoma.

Candida species are a normal commensal of the oral cavity in healthy individuals, but can become an opportunistic pathogen when the oral ecosystem is unbalanced. Several virulence attributes have been identified in candidal infection, among which are the hydrolases, including the secreted aspartyl proteinases (Saps). This study evaluated and compared the in vitro level of Saps from Candida albicans in nonsmokers, smokers, and patients with leukoplakia and oral squamous cell carcinoma (OSCC). Candida cell count (CCC) at 48 h was also assessed. The Sap level was measured by spectrophotometry in 38 clinical isolates of C. albicans obtained from the oral cavity of the four different groups. Culturing was done in yeast carbon base-bovine serum albumin. Speciation of Candida was performed by using a Candida identification kit, and CCC was measured by hemocytometer. Sap levels and CCC were higher in individuals with leukoplakia and OSCC than in nonsmokers or smokers (P = 0.001); however, there was no significant difference in Sap levels or CCC between smokers and nonsmokers (P = 0.529). Further, an intragroup correlation between CCC and Sap level was also observed. The higher level of Saps from C. albicans in individuals with leukoplakia and OSCC suggests that this pathogen plays a role in disease development and could aid in identifying the pathogenic commensal.

Estimation of salivary protein thiols and total antioxidant power of saliva in brain tumor patients.

BACKGROUND: The role of oxidative stress in the genesis of various types of cancers is well established. Several chemical, cell culture and animal studies also indicate that antioxidants may slow or even prevent the development of cancer. Brain is considered abnormally sensitive to oxidative damage as brain tissue has high rate of oxygen consumption, high lipid content and relatively low antioxidant defenses, compared to other tissues. MATERIALS AND METHODS: The study design chosen for the present study was cross sectional. The biochemical parameters that were estimated in saliva manually using spectrophotometric methods were ferric reducing antioxidant power (FRAP) assay--a direct measure of total antioxidant activity of biological fluids and protein thiols. The physical parameters of saliva that were also assessed were salivary flow rate, pH of the saliva and the osmolality of the saliva. RESULTS: The mean values of salivary flow rate and pH were significantly decreased among malignant and benign tumor patients whereas the salivary osmolality was significantly increased in both the groups of patients. The mean values of salivary FRAP were significantly reduced among malignant and benign tumor patients. However, the salivary protein thiols were significantly increased in these patients. CONCLUSION: Hence with these observations it can be concluded that in saliva, besides the physical characteristics, salivary FRAP and protein thiol levels are appropriate indicators of the antioxidant status in brain tumor patients.

Assessment of serum L-fucose in brain tumor cases.

BACKGROUND: Glycosylation of altered tumor cell in relation to cellular heterogeneity in human intracranial tumors remains relatively unexposed. Serum protein-bound carbohydrate, L-Fucose is reported to be overexpressed during tumor progression by many investigators. Therefore, there is a need to determine the diagnostic, prognostic, functional significance of glycoprotein elevations in various cases of tumors. OBJECTIVE: The objective of the present study was to evaluate the clinical utility of serum L-fucose in patients with brain tumor. MATERIALS AND METHODS: Serum glyco-conjugate levels were estimated in 99 patients with brain tumors. Estimation of L-fucose was carried out colorimetrically by the method of Winzler using cysteine hydrochloride. RESULTS: There was a significant increase in L-fucose level in most of the patients. In the posttreatment cases, the L-fucose levels were apparently low compared to preoperative values. CONCLUSION: Our results showed that the rise in serum L-fucose may be used as a general marker for brain tumors in addition to other markers.

Antioxidants and lipid peroxidation status in diabetic patients with and without complications.

BACKGROUND: Oxidative stress is involved in the pathophysiology of diabetes mellitus. METHODS: In the present study, 68 patients with type 2 diabetes mellitus and 31 clinically healthy individuals were evaluated. The patients were divided into two groups. Group 1 included 29 patients without diabetic complications and group 2 consisted of 39 patients with diabetic complications. Erythrocyte glutathione, superoxide dismutase, and thiobarbituric acid-reactive substance levels as well as plasma antioxidant vitamins C and E, and serum total glutathione-S-transferase, ceruloplasmin, and protein thiols were estimated by using spectro-photometer. RESULTS: A significant decrease of erythrocyte glutathione was observed in group 1 when compared with the controls. Thiols decreased in group 2. An increase in glutathione-S-transferase, ceruloplasmin, superoxide dismutase, and vitamins C and E levels was noted in patients with diabetes mellitus. Thiobarbituric acid-reactive substance levels decreased in group 1 but increased in group 2 when compared with the controls. CONCLUSION: In the present study, tendency of most of the antioxidants to rise in diabetes could probably be due to an adaptive response to the pro-oxidant milieu of the diabetic state. Hence, we suggest that supplementation with dietary antioxidants especially antioxidant vitamins accompanied by change in lifestyle might help to reduce damage brought about by free radical toxicity in diabetes mellitus.

Assessment of total sialic acid and lipid-bound sialic acid in management of brain tumors.

BACKGROUND: Glycoconjugate molecules expressed at the plasma membrane of mammalian cells have been reported to be associated with tumor progression. The measurement of total sialic acid (TSA) and lipid-bound sialic acid (LBSA) in the cerebrospinal fluid (CSF) is suggested to be useful for the diagnosis of brain tumors. But there are very few reports available on the serum glycoconjugate levels in patients with brain tumors. OBJECTIVE: The objective of this study is to check the feasibility of using serum glycoconjugates such as TSA and LBSA as tumor markers in brain tumor patients. MATERIALS AND METHODS: Colorimetric estimation of TSA using diphenylamine was done on 100 patients with intracranial tumors; follow-up study was carried out in 24 cases. The LBSA fraction was isolated from the serum of 68 brain tumor patients and evaluated using phosphotungstic acid and resorcinol; follow-up study was done on 23 patients. The various types of brain tumors included in this study were glioma, meningioma, and acoustic neurinoma as well as some other types such as medulloblastoma, secondary tumors, and craniopharyngioma. RESULTS: There was no significant difference between the TSA and LBSA concentrations seen in pretreatment or post-treatment cases and that seen in control subjects. DISCUSSION: TSA and LBSA do not have the ability to discriminate between benign and malignant brain tumors. TSA and LBSA appear to be tumor markers of very limited value in patients with brain tumors.

Plasma protein oxidation in patients with brain tumors.

OBJECTIVE: Proteins can undergo numerous covalent changes on exposure to oxidants. Oxidative modification of protein in vivo may affect a variety of cellular functions. Protein oxidation in vivo is a natural consequence of aerobic life. Oxygen radicals and other activated oxygen species generated as byproducts of cellular metabolism or from environmental sources cause modifications to the amino acids of proteins that generally result in loss of protein function/enzymatic activity. It is now well known that reactive oxygen species (ROS) play a key role in human cancer development. Moreover, the brain is especially vulnerable to ROS mediated injury. METHOD: Therefore, in the present study, protein oxidation was assessed in the plasma of 17 patients with brain tumors and 16 age and gender-matched controls by measuring protein thiols and protein carbonyls spectrophotometrically. RESULTS: There was a significant decrease in protein thiols and carbonyls in malignant cases of brain tumors when compared with the control group. No significant change in protein thiols was noted in benign cases compared to controls. A comparison of levels in benign and malignant cases for both the parameters also showed no significant difference. DISCUSSION: Thus, free radical toxicity does lead to protein oxidation in patients with brain tumors.

Relationship between markers of lipid peroxidation, thiol oxidation and Glasgow coma scale scores of moderate head injury patients in the 7 day post-traumatic period.

OBJECTIVE: Epidemiologic works reveal that moderate head injury (MHI) is more frequent and a substantial number of these patients develop complications resulting in neurological disabilities. Reactive oxygen species (ROS) play a major role in post-traumatic neuronal damage following traumatic head injury. Thus, the current study analysed the post-traumatic changes in the erythrocyte markers of oxidative damage and the relationship between these parameters and Glasgow coma scale (GCS) scores of MHI patients during the 7 day study period. METHODS: Peripheral venous blood samples were taken at the time of hospital admission (d1 of injury) and on d7 from 25 MHI patients (admission GCS score > 8). These were compared with samples from 25 healthy individuals (normal controls, NC). GCS scores were recorded at the same time points of the study period. Erythrocyte lipid peroxidation (LP) and thiol oxidation levels were estimated and compared with that of NC. The relationship between GCS scores and erythrocyte markers were also studied. RESULTS: Erythrocyte thiobarbituric acid reactive substance (TBARS) levels reflecting lipid peroxidative damage remained significantly elevated at both time points of the study period in MHI patients as compared to NC (p < 0.001 ). There was a significant decrease in the level of nonprotein thiols in MHI patients as compared to NC (p < 0.01) at the same time points of the study. However, on d7 there were no further significant changes in the markers of oxidative damage in MHI patients as compared to on d1. CONCLUSION: These findings suggest that a condition of oxidative stress occurs during the entire post-traumatic period in MHI patients and the utility of markers of oxidative damage in the prognosis of head injury needs to be addressed in further works.

Susceptibility of peripheral lymphocytes of brain tumour patients to in vitro radiation-induced DNA damage, a preliminary study.

OBJECTIVE: The present investigation aimed to study the susceptibility of lymphocytes collected from brain tumour patients to radiation-induced DNA damage under in vitro conditions. METHODS: The peripheral lymphocytes collected from brain tumour patients were exposed to 2-Gy gamma radiation. Susceptibility of lymphocytes to radiation-induced DNA damage and their repair ability was assessed by alkaline comet assay. RESULTS: Lymphocytes of patients with benign and malignant tumour had a significantly higher (p < 0.001) baseline DNA damage compared to lymphocytes from normal subjects. A significant increase (p < 0.001) in DNA damage was observed immediately after irradiation of lymphocytes from healthy subjects and brain tumour patients. However, at 1 h after irradiation the level of DNA damage dropped significantly (p < 0.001) compared to that of immediately after irradiation of respective groups. DISCUSSION: In conclusion, the lymphocytes of brain tumour patients possess a higher level of basal DNA damage and exhibit a higher susceptibility to a clastogenic agent like radiation.