Cell cycle inhibitors, such as the cyclin-dependent kinase (Cdk) inhibitor proteins and retinoblastoma (Rb) family members, control exit from cell during development of a variety terminally differentiated tissues. It is unclear whether sustained expression these required to prevent re-entry in quiescent cells. The organ Corti (cochlear sensory epithelium) pars intermedia (intermediate lobe pituitary) are two tissues that share characteristic ongoing division mice lacking either p27(Kip1) Cdk...

Fine particulate air pollution (PM2.5) is a global health concern, as exposure to PM2.5 has consistently been found be associated with increased cardiovascular morbidity and mortality. Although adult traffic related PM2.5, which largely derived from diesel exhaust (DE), cardiac hypertrophy, there are limited investigations into the potential effect of in utero early life on susceptibility heart disease. In this study, we investigate DE failure.Female C57BL/6 J mice were exposed either...

The R47H variant in the microglial TREM2 receptor is a strong risk factor for Alzheimer's disease (AD). To characterize processes affected by we performed integrative network analysis of genes expressed brains AD patients with R47H, sporadic without and polycystic lipomembranous osteodysplasia sclerosing leukoencephalopathy (PLOSL), systemic early onset dementia caused loss-of function mutations or its adaptor TYROBP. While had few perturbed immune genes, demonstrated upregulation interferon...

Missense mutations in myosin heavy chain 7 (MYH7) are a common cause of hypertrophic cardiomyopathy (HCM), but the molecular mechanisms underlying MYH7-based HCM remain unclear. In this work, we generated cardiomyocytes derived from isogenic human induced pluripotent stem cells to model heterozygous pathogenic MYH7 missense variant, E848G, which is associated with left ventricular hypertrophy and adult-onset systolic dysfunction. MYH7E848G/+ increased cardiomyocyte size reduced maximum...

The effects of human papillomavirus type 18 (HPV-18) E6 and E7 proteins on global patterns host gene expression in primary keratinocytes grown organotypic raft culture system were assessed. Primary infected with retroviruses that express the wild-type HPV-18 genes from native differentiation-dependent HPV enhancer-promoter. Total RNA was isolated cultures used to generate probes for querying Affymetrix U95A microarrays, which contain >12,500 sequences. Quadruplicate arrays each...

Abstract Background The response of mammalian glial cells to chronic degeneration and trauma is hypothesized be incompatible with support neuronal function in the central nervous system (CNS) retina. To test this hypothesis, we developed an inducible model proliferative reactive gliosis absence degenerative stimuli by genetically inactivating cyclin-dependent kinase inhibitor p27 Kip1 ( or Cdkn1b ) adult mouse determined outcome on retinal structure function. Results p27-deficient Müller...

Abstract Triggering receptor expressed on myeloid cells 2 (TREM2) is an immunoglobulin-like by certain cells, such as macrophages, dendritic osteoclasts, and microglia. In the brain, TREM2 plays important role in immune function of microglia, its dysfunction linked to various neurodegenerative conditions humans. Ablation or adaptor protein TYROBP causes polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy (also known Nasu-Hakola disorder) early onset dementia, whereas...

ABSTRACT The productive program of human papillomaviruses (HPVs) in epithelia is tightly linked to squamous differentiation. E7 proteins high-risk HPV genotypes efficiently inactivate the pRB family that control cell cycle, triggering S phase suprabasal keratinocytes. This ability has until now not been demonstrated for low-risk HPV-6 or HPV-11 proteins. An inducible system which HPV-16 fused ligand binding domain estrogen receptor (ER) was described by Smith-McCune et al. (K. Smith-McCune,...

Atrial natriuretic factor (ANF) is abundantly expressed in atrial cardiomyocytes throughout ontogeny and ventricular the developing heart. However, during cardiac failure hypertrophy, ANF expression can reappear adult cardiomyocytes. The transcription Nkx2-5 one of major transactivators gene We identified binding at three 5′ regulatory elements (kb −34, −31, −21) proximal promoter by ChIP assay using neonatal mouse 3C analysis revealed close proximity between distal region. A 5.8-kb fragment...

ABSTRACT The human papillomavirus (HPV) E7 protein promotes S-phase reentry in a fraction of postmitotic, differentiated keratinocytes. Here we report that these cells contain an inherent mechanism opposes E7-induced DNA replication. In organotypic raft cultures primary keratinocytes, neither cyclin E nor p21cip1 is detectable situ. However, E7-transduced not S phase accumulate abundant and p21cip1. We show normally rapidly degraded by proteasomes. the presence or E6/E7, p21cip1, E, E2...

Loss of the cyclin-dependent kinase inhibitor p27 Kip1 leads to an overall increase in animal growth, pituitary tumors, and hyperplasia hematopoietic organs, yet it is unknown whether all cells function autonomously response activity or certain take cues from their neighbors. In addition, there currently no genetic evidence that tumor suppression by cell-autonomous because biallelic gene inactivation absent tumors arising hemizygous mice. We have addressed these questions with...

In utero exposure to diesel exhaust air pollution has been associated with increased adult susceptibility heart failure in mice, but the mechanisms by which this promotes are poorly understood. To identify potential transcriptional effects that mediate susceptibility, we have performed RNA sequencing analysis on hearts from mice were exposed and subsequently undergone transverse aortic constriction. We identified 3 target genes, Mir133a-2, Ptprf, Pamr1, demonstrate dysregulation after...

Developmental exposure to particulate matter air pollution is harmful cardiovascular health, but the mechanisms by which this mediates susceptibility heart disease poorly understood. We have previously shown, in a mouse model, that gestational diesel exhaust (DE) results increased cardiac hypertrophy, fibrosis and failure adult offspring following transverse aortic constriction. In study, we analyzed gene expression neonatal cardiomyocytes after RNA-sequencing identified 300 genes are...

CHF1/Hey2 is a Notch-responsive basic helix–loop-helix transcription factor involved in cardiac development. Common variants Hey2 are associated with Brugada syndrome. We hypothesized that absence of would result abnormal cellular electrical activity, altered conduction system (CCS) development, and increased arrhythmogenesis. isolated neonatal CHF/Hey2-knockout (KO) myocytes measured action potentials ion channel subunit gene expression. also crossed myocardial-specific CHF1/Hey2-KO mice...

AbstractMice lacking the p27Kip1 Cdk inhibitor, like mice Rb, develop pituitary tumors involving pars intermedia melanotrophs, yet are genetically distinct from Rb derived as they exhibit haploid insufficiency. We compared with constitutive and tissue specific null mutations to arising in knockout aim of determining whether distinguished by quantitative or qualitative differences. The rate tumor development was strongly influenced strain background due polygenic modifiers C57BL/6J versus...

Generating cardiomyocytes from embryonic stem cells is an important technique for understanding cardiovascular development, the origins of diseases and also providing potential reagents cardiac repair. Numerous methods have been published but often are technically challenging, complex, not easily adapted to assessment specific gene contributions myocyte differentiation. Here we report development optimized protocol induce differentiation mouse myocytes that simplified genetic studies....

The tumor-suppressor gene p53 may transactivate the transcription of genes that down-regulate cellular growth-related and become oncogenic as a result production mutant proteins or loss its protein expression. This study reports alterations highly conserved consensus intervening sequences at splice junctions lead to inactivation gene. Analyses with combined polymerase chain reaction single-strand conformational polymorphism direct DNA sequencing DNAs amplified by means asymmetric...

Abstract Background Strong associations have been observed between exposure to fine ambient particulate matter (PM 2.5 ) and adverse cardiovascular outcomes. In particular, traffic related PM has associated with increases in left ventricular hypertrophy, a strong risk factor for mortality. As much of is derived from diesel exhaust (DE), we investigated the effects chronic DE on cardiac hypertrophy heart failure adult mouse by exposing mice combined either two models hypertrophy: angiotensin...

Standard transgenic cell line generation requires screening 100-1000s of colonies to isolate correctly edited cells. We describe CRISPRa On-Target Editing Retrieval (CRaTER) which enriches for cells with on-target knock-in a cDNA-fluorescent reporter transgene by transient activation the targeted locus followed flow sorting recover show CRaTER recovers rare heterozygous, biallelic-editing transcriptionally-inactive MYH7 in human induced pluripotent stem (hiPSCs), enriching on average 25-fold...

ABSTRACT The human papillomavirus type 18 (HPV-18) E7 protein promotes S-phase reentry in postmitotic, differentiated keratinocytes squamous epithelium to facilitate vegetative viral DNA amplification. To examine the nature and fate of cells that reenter S phase, organotypic cultures primary transduced with HPV-18 were pulse-chase-pulse-labeled 3 H-thymidine ( H-TdR) bromodeoxyuridine (BrdU). kinetics appearance doubly labeled suprabasal demonstrate expression did not promote prolonged...