Gregariousness in the animal world.

Thesis (M.A.)--Boston Universit

ERK Activation Requires CaM Kinases in MCF-7 Breast Cancer Cells

Abstract A key signaling pathway involved in regulating cell growth and proliferation throughout the body is the ERK signaling pathway. ERK is activated via numerous pathways including intracellular calcium release downstream of G-Protein Coupled Receptors (GPCRs). Carbachol, a GPCR-agonist, both increases intracellular calcium and ERK activation in MCF-7 cells. ERK activation and control of cell growth may act through the transcription factor Elk-1. Our goal was to elucidate the specific proteins and kinases upstream of ERK in MCF-7 cells treated with carbachol. Secondly, we wanted to investigate the potential involvement of Elk-1 downstream of ERK. Carbachol treatment of MCF-7 cells triggered ERK and Elk-1 phosphorylation within 5 minutes. Interestingly, transfection of cells with shRNAs directed to either CaM KKα or CaM KIγ significantly inhibited carbachol activation of ERK. Phosphorylation of Elk-1, following carbachol stimulation, was also blocked in siERK2 transfected cells, suggesting that ERK2 is required for carbachol\u27s activation of Elk-1. Our results suggest that carbachol treatment of MCF-7 cells activates ERK and cell growth through CaM KK, CaMKI, and an Elk-1-dependent pathway

M3-Muscarinic Receptor Activation of ERK and Cell Growth Requires Calcium/Calmodulin-dependent Protein Kinases in MCF-7 Cells

Abstract The extracellular signal-regulated protein kinase (ERK) signaling pathway is found in diverse cells throughout the human body. ERK activation has been implicated in breast cancer cell growth and proliferation. Studies have shown that ERK is activated by carbachol, a G Protein-Coupled Receptor (GPCR) agonist, which increases intracellular calcium in MCF-7 cells. The calcium/calmodulin-dependent protein kinase (CaM K) family of proteins including CaM KK, CaM KI, and CaM KII can be activated by increased intracellular calcium. Our goal was to determine whether CaM Ks may be responsible for ERK activation and cell proliferation in carbachol-treated MCF-7 cells and evaluate which GPCR was responsible for these events. Carbachol treatment of MCF-7 cells triggered ERK 1/2 phosphorylation within 5 minutes. Treatment with KN-93, a general CaM Kinase inhibitor and the MEK inhibitor U0126 blocked ERK activation. Carbachol increased MCF-7 cell growth nearly 4-fold, an effect that was also dependent upon CaM Ks and MEK. Interestingly, CaM KK was responsible for ERK activation and cell growth. Pretreatment of MCF-7 cells with 4-DAMP, a selective M3 receptor antagonist, completely blocked carbachol’s activation of ERK and cell growth. Taken together these results suggest that carbachol stimulated ERK phosphorylation and MCF-7 cell growth by the M3 subtype GPCR receptor perhaps through CaM KK

Implications of Co-Residential Status for Parenting Programs Targeting Adolescent Mothers

When a teenager becomes a parent, what type of family living arrangements best support her parenting competence? Conventional wisdom suggests that the mother\u27s family of origin would be the best arrangement, but contemporary adolescents do their parenting in a variety of situations. Results from the study reported here, examining mothers living in three different types of arrangements, suggest that living with a parenting partner of the same generational age as the mother supports more positive parenting attitudes. A strengths-based, youth development approach to parenting education for adolescents is discussed in which the interpersonal context of adolescent parenting is explicitly addressed

ERK Activation and Cell Growth Require CaM Kinases in MCF-7 Breast Cancer Cells

Previous studies on MCF-7 breast cancer cells have shown that the G-protein coupled receptor (GPCR) agonist carbachol increases intracellular calcium levels and the activation of extracellular signal-regulated kinase (ERK). Calcium and calmodulin regulate the calcium/calmodulin- dependent kinase (CaM kinase) family of proteins that have been proposed to regulate ERK and gene transcription. Our results suggest that both estrogen (E2) and carbachol treatment of MCF-7 breast cancer cells trigger phosphorylation of ERK I /2 and the transcription factor Elk-1. Carbachol and estrogen triggered nearly a four- to sixfold increase in MCF-7 cell proliferation by 96 h, respectively. Carbachol-stimulated ERK activation and cell growth was completely blocked by the Muscarinic M3- subtype GPCR inhibitor, 4-DAMP, and siRNA against the M3-subtype GPCR. Interestingly, blockade of CaM KK with the selective inhibitor ST0-609 prevented carbachol activation CaM KI, ERK, Elk-1 , and cell gro\vth. Consistent with these observations, knockdown of CaM KKa and CaM Kly with shRNA-containing plas1nids blocked ERK activation by carbachol. In addition, Elk-I phosphorylation and luciferase activity in response to carbachol treat1nent was also dependent upon CaM kinases and was inhibited by U0126, ST0-609, and siRNA knockdown of CaM kinases and ERK2. Finally, blockade of either CaM KK (with ST0-609) or ERK (with U0126) activities resulted in the inhibition of carbachol- and estrogen-mediated cyclin Dl expression and MCF-7 cell growth. Taken together, our results suggest that carbachol treatment of MCF-7 cells activates CaM KI, ERK, the transcription factor Elk-1 , cyclin D 1, and cell grovvth through CaM KK

Psychosocial factors and cancer incidence (PSY-CA):Protocol for individual participant data meta-analyses

OBJECTIVES: Psychosocial factors have been hypothesized to increase the risk of cancer. This study aims (1) to test whether psychosocial factors (depression, anxiety, recent loss events, subjective social support, relationship status, general distress, and neuroticism) are associated with the incidence of any cancer (any, breast, lung, prostate, colorectal, smoking-related, and alcohol-related); (2) to test the interaction between psychosocial factors and factors related to cancer risk (smoking, alcohol use, weight, physical activity, sedentary behavior, sleep, age, sex, education, hormone replacement therapy, and menopausal status) with regard to the incidence of cancer; and (3) to test the mediating role of health behaviors (smoking, alcohol use, weight, physical activity, sedentary behavior, and sleep) in the relationship between psychosocial factors and the incidence of cancer.METHODS: The psychosocial factors and cancer incidence (PSY-CA) consortium was established involving experts in the field of (psycho-)oncology, methodology, and epidemiology. Using data collected in 18 cohorts (N = 617,355), a preplanned two-stage individual participant data (IPD) meta-analysis is proposed. Standardized analyses will be conducted on harmonized datasets for each cohort (stage 1), and meta-analyses will be performed on the risk estimates (stage 2).CONCLUSION: PSY-CA aims to elucidate the relationship between psychosocial factors and cancer risk by addressing several shortcomings of prior meta-analyses.</p

Genetic effects on gene expression across human tissues

Characterization of the molecular function of the human genome and its variation across individuals is essential for identifying the cellular mechanisms that underlie human genetic traits and diseases. The Genotype-Tissue Expression (GTEx) project aims to characterize variation in gene expression levels across individuals and diverse tissues of the human body, many of which are not easily accessible. Here we describe genetic effects on gene expression levels across 44 human tissues. We find that local genetic variation affects gene expression levels for the majority of genes, and we further identify inter-chromosomal genetic effects for 93 genes and 112 loci. On the basis of the identified genetic effects, we characterize patterns of tissue specificity, compare local and distal effects, and evaluate the functional properties of the genetic effects. We also demonstrate that multi-tissue, multi-individual data can be used to identify genes and pathways affected by human disease-associated variation, enabling a mechanistic interpretation of gene regulation and the genetic basis of diseas

Depression, anxiety, and the risk of cancer: An individual participant data meta-analysis

BACKGROUND: Depression and anxiety have long been hypothesized to be related to an increased cancer risk. Despite the great amount of research that has been conducted, findings are inconclusive. To provide a stronger basis for addressing the associations between depression, anxiety, and the incidence of various cancer types (overall, breast, lung, prostate, colorectal, alcohol-related, and smoking-related cancers), individual participant data (IPD) meta-analyses were performed within the Psychosocial Factors and Cancer Incidence (PSY-CA) consortium. METHODS: The PSY-CA consortium includes data from 18 cohorts with measures of depression or anxiety (up to N = 319,613; cancer incidences, 25,803; person-years of follow-up, 3,254,714). Both symptoms and a diagnosis of depression and anxiety were examined as predictors of future cancer risk. Two-stage IPD meta-analyses were run, first by using Cox regression models in each cohort (stage 1), and then by aggregating the results in random-effects meta-analyses (stage 2). RESULTS: No associations were found between depression or anxiety and overall, breast, prostate, colorectal, and alcohol-related cancers. Depression and anxiety (symptoms and diagnoses) were associated with the incidence of lung cancer and smoking-related cancers (hazard ratios [HRs], 1.06-1.60). However, these associations were substantially attenuated when additionally adjusting for known risk factors including smoking, alcohol use, and body mass index (HRs, 1.04-1.23). CONCLUSIONS: Depression and anxiety are not related to increased risk for most cancer outcomes, except for lung and smoking-related cancers. This study shows that key covariates are likely to explain the relationship between depression, anxiety, and lung and smoking-related cancers. PREREGISTRATION NUMBER: https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=157677

Genetic effects on gene expression across human tissues

Characterization of the molecular function of the human genome and its variation across individuals is essential for identifying the cellular mechanisms that underlie human genetic traits and diseases. The Genotype-Tissue Expression (GTEx) project aims to characterize variation in gene expression levels across individuals and diverse tissues of the human body, many of which are not easily accessible. Here we describe genetic effects on gene expression levels across 44 human tissues. We find that local genetic variation affects gene expression levels for the majority of genes, and we further identify inter-chromosomal genetic effects for 93 genes and 112 loci. On the basis of the identified genetic effects, we characterize patterns of tissue specificity, compare local and distal effects, and evaluate the functional properties of the genetic effects. We also demonstrate that multi-tissue, multi-individual data can be used to identify genes and pathways affected by human disease-associated variation, enabling a mechanistic interpretation of gene regulation and the genetic basis of disease

Genomic epidemiology of SARS-CoV-2 in a UK university identifies dynamics of transmission

AbstractUnderstanding SARS-CoV-2 transmission in higher education settings is important to limit spread between students, and into at-risk populations. In this study, we sequenced 482 SARS-CoV-2 isolates from the University of Cambridge from 5 October to 6 December 2020. We perform a detailed phylogenetic comparison with 972 isolates from the surrounding community, complemented with epidemiological and contact tracing data, to determine transmission dynamics. We observe limited viral introductions into the university; the majority of student cases were linked to a single genetic cluster, likely following social gatherings at a venue outside the university. We identify considerable onward transmission associated with student accommodation and courses; this was effectively contained using local infection control measures and following a national lockdown. Transmission clusters were largely segregated within the university or the community. Our study highlights key determinants of SARS-CoV-2 transmission and effective interventions in a higher education setting that will inform public health policy during pandemics.</jats:p