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Biology subjects

Sussman, M.

Publications and source records attributed to Sussman, M..

9 recordsLinked to original sources

Vape Aerosol Exposure Decreases Voluntary Exercise in Mice

Electronic cigarette use (vaping) is generally accepted as a less harmful alternative to traditional cigarette smoking. However, so called "never smokers" including young adults and teenagers are often recruited through targeted advertising and appealing vape juice flavors. Any fitness cost of vaping matters for young people who exercise or play sports. We therefore tracked running distance in a mouse model of voluntary exercise to assess the effect of vaping. Specifically, we asked whether vaping decreases exercise performance, and whether there is progressive change over time. Experimental groups of mice subjected to whole-body inhalation exposure and corresponding non-exposed controls were housed in cages with running wheels. Daily wheel laps were recorded over the 8-week exposure period and analyzed using hierarchical generalized additive mixed models. Vape-exposed females ran 32% fewer laps than control females. Vape-exposed males showed a smaller reduction of 15% that did not reach statistical significance. The reduction was 10,025 laps per day larger in females than in males (p = 0.02). In females, the difference was present from the first weeks of exposure and was larger by the end of the study. The results demonstrate that vape exposure is associated with reduced voluntary aerobic activity. The analysis illustrates how nonlinear longitudinal modeling clarifies both the size and the temporal character of treatment effect in this experimental model. These findings reveal that nicotine-free components of vape aerosol reduce voluntary aerobic activity, highlighting an overlooked facet of vaping toxicity research which is particularly relevant for young and physically active users.

pharmacology and toxicology↗

Lipid Transfer Proteins and PI4KIIα Generate a Phosphoinositide-Linked Proteome

Phosphoinositide (PIPn) lipid second messengers in membranes regulate a myriad of cellular processes. In the cytosol, the phosphatidylinositol (PI) 3-kinase (PI3K)/Akt pathway is scaffolded on IQGAP1 to facilitate the activation of Akt by the synthesis of PI3,4,5P3. In the nucleus, PIPn signaling occurs in regions devoid of membranes via their stable association with proteins. While several of these proteins have been identified, understanding the extent and impact of protein-linked PIPn signaling warrants further investigation. The tumor suppressor p53, was shown in the companion paper to be regulated by PI transfer proteins (PITPs) and a PI 4-kinase (PI4KII), which are required to form p53-PIPn complexes that assemble a nuclear PI3K/Akt pathway. Here we report that class I PITPs (PITP/{beta}) and PI 4-kinase initiate PIPn linkages to many different proteins. PITP/{beta} and PI4KII accumulate in the nucleoplasm in response to stress and are necessary to synthesize nuclear PIPns linked to proteins. These PITP/{beta}-dependent protein-PIPn complexes are detected by metabolically labeling cells with the PIPn precursor [H3]-myo-inositol and resist denaturation and SDS-PAGE, indicating that these protein-PIPn complexes represent a putative posttranslational modification. Proteomic analyses of proteins that are regulated by PITP/{beta} and/or are linked to PI4,5P2 have identified an emerging PIPylome that is enriched in metabolic, signaling, cytoskeletal and DNA repair pathway components. Taken together, these data provide evidence for an emerging proteome with linked PIPns that represent a PIPn signaling paradigm that is distinct from the membrane-localized pathway but utilizes many of the same PIP kinases and phosphatases. In briefPhosphatidylinositol transfer proteins and PI 4-kinase initiate a PIPn-linked protein network in membrane-free regions.

cell biology↗

The PIDDosome controls cardiomyocyte polyploidization during postnatal heart development

The adult mammalian heart is characterized by post-mitotic polyploid cardiomyocytes (CMs). Understanding how CMs regulate cell cycle exit and ploidy can help developing new heart regenerative therapies. Here, we uncover that the PIDDosome, a multi-protein complex activating the endopeptidase Caspase-2, helps to implement a CM-specific differentiation program that limits ploidy during postnatal heart development. DNA content analyses show that PIDDosome-loss causes a cell-autonomous increase in nuclear and cellular CM ploidy. Remarkably, increased ploidy does not affect cardiac structure nor function. PIDDosome-imposed ploidy restriction commences at postnatal day 7 (P7), reaching a plateau on P14. PIDDosome activation requires ANKRD26, targeting PIDD1 to mother centrioles. Opposite to prior observations in liver development, the PIDDosome limits CM polyploidization in a p53-independent manner but reliant on p21/Cdkn1a, a notion supported by nuclear RNA sequencing and genetic deletion experiments. Our results provide new insights how proliferation of polyploid CMs is restricted during postnatal heart development.

developmental biology↗

Cardiac p16 Expression Following Vape Exposure with Nicotine Shows Sex-Specific Induction in Males but not Females

Vaping is marketed as a safe alternative to traditional cigarette smoking, but multiple studies demonstrate deleterious cardiopulmonary effects including cardiac function decline and fibrotic remodeling with alveolar size enlargement. Nicotine, a common constituent of vaping aerosol, stimulates p16 expression in pulmonary tissue but the impact on cardiac tissue remains unclear. In this study, mice were exposed to e-cigarette vape aerosol either containing nicotine (Vape Nicotine; VN) or without nicotine (Vape 0; V0). Non-exposed (No Vape; NoV) mice were used as controls. Cardiac effects were assessed by echocardiography, histology, and immunofluorescence to determine changes in function, morphology, p16, and Discoidin Domain Receptor 2 (DDR2). VN depressed cardiac function and increased collagen deposition relative to V0 and NoV. Interestingly, p16 expression was increased in cardiomyocytes and interstitial cells of male mice while remaining unchanged in females. In contrast to VN, V0 had no significant impact on cardiac function or p16 expression in males. Furthermore, collagen deposition in the V0 group was significantly lower than the VN group. Subsequent cardiac fibroblast analysis using DDR2 revealed increased expression within the V0 group relative to VN and NoV. Collectively, these findings show collagen accumulation as well as p16 expression prompted by vaping is mediated by nicotine as a constituent of vape juice. In contrast, vape aerosol alone promotes accrual of cardiac fibroblasts without concomitant changes in collagen accumulation or p16 expression. These results are the first to identify p16 induction with pathologic collagen deposition by exposure to vape aerosol containing nicotine in male cardiac tissue. The underlying basis for sex-specific differences in cardiac responses to vape aerosol exposure warrant further investigation, particularly those involving cellular and molecular changes that may lead to pathologic changes later in life.

molecular biology↗

Myocardial infarction injury is exacerbated by nicotine in vape aerosol exposure

RationaleVaping is touted as a safer alternative to traditional cigarette smoking but the full spectrum of harm reduction versus comparable risk remains unresolved. Elevated bioavailability of nicotine in vape aerosol together with known risks of nicotine exposure may result in previously uncharacterized cardiovascular consequences of vaping. ObjectiveAssess the impact of nicotine exposure via vape aerosol inhalation upon myocardial response to infarction injury. Methods and ResultsFlavored vape juice containing nicotine (5 mg / ml) or vehicle alone (0 mg) was delivered using identical 4-week treatment protocols. Mice were subjected to acute myocardial infarction injury and evaluated for outcomes of cardiac structure and function. Findings reveal that nicotine exposure leads to worse outcomes with respect to contractile performance regardless of sex. Non-myocyte interstitial cell accumulation following infarction significantly increased with exposure to vape aerosol alone, but a comparable increase was not present when nicotine was included. ConclusionsMyocardial function after infarction is significantly decreased after exposure to nicotine vape aerosol irrespective of sex. Comparable loss of contractile function was not observed in mice exposed to vape aerosol alone, highlighting the essential role of nicotine in loss of contractile function. Increased vimentin immunoreactivity was observed in the vape alone group compared to control and vape nicotine. The correlation between vaping, interstitial cell responses, and cardiac remodeling leading to impaired contractility warrants further investigation. Public health experts seeking to reduce vaping-related health risks should consider messaging that highlights the increased cardiovascular risk especially with nicotine-containing aerosols.

molecular biology↗

Human CardioChimeras: Creation of a Novel \"Next Generation\" Cardiac Cell

BackgroundCardioChimeras (CCs) produced by fusion of murine c-kit+ cardiac interstitial cells (cCIC) with mesenchymal stem cells (MSCs) promote superior structural and functional recovery in a mouse model of myocardial infarction (MI) compared to either precursor cell alone or in combination. Creation of human CardioChimeras (hCC) represents the next step in translational development of this novel cell type, but new challenges arise when working with cCICs isolated and expanded from human heart tissue samples. The objective of the study was to establish a reliable cell fusion protocol for consistent optimized creation of hCCs and characterize fundamental hCC properties.\n\nMethods and ResultsCell fusion was induced by incubating human cCICs and MSCs at a 2:1 ratio with inactivated Sendai virus. Hybrid cells were sorted into 96-well microplates for clonal expansion to derive unique cloned hCCs, which were then characterized for various cellular and molecular properties. hCCs exhibited enhanced survival relative to the parent cells and promoted cardiomyocyte survival in response to serum deprivation in vitro.\n\nConclusionsThe generation of hCC is demonstrated and validated in this study, representing the next step toward implementation of a novel cell product for therapeutic development. Feasibility of creating human hybrid cells prompts consideration of multiple possibilities to create novel chimeric cells derived from cells with desirable traits to promote healing in pathologically damaged myocardium.\n\nClinical PerspectiveO_LI\"Next generation\" cell therapeutics will build upon initial findings that demonstrate enhanced reparative action of combining distinct cell types for treatment of cardiomyopathic injury.\nC_LIO_LIDifferential biological properties of various cell types are challenging for optimization of delivery, engraftment, persistence, and synergistic action when used in combination.\nC_LIO_LICreation of a novel hybrid cell called a CardioChimera overcomes limitations inherent to use of multiple cell types.\nC_LIO_LICardioChimeras exhibit unique properties relative to either parental cell anticipated to be advantageous in cellular therapeutic applications.\nC_LIO_LICardioChimeras have now been created and characterized using cells derived from human heart tissue, advancing initial proof of concept previously demonstrated with mice.\nC_LIO_LICardioChimeras represent an engineered solution that can be implemented as a path forward for improving the outcome of myocardial cell therapy.\nC_LI

cell biology↗

Enhancing Myocardial Repair with CardioClusters

BackgroundCellular therapy to treat heart failure is an ongoing focus of intense research and development, but progress has been frustratingly slow due to limitations of current approaches. Engineered augmentation of established cellular effectors overcomes impediments, enhancing reparative activity with improved outcomes relative to conventional techniques. Such next generation implementation includes delivery of combinatorial cell populations exerting synergistic effects. Concurrent isolation and expansion of three distinct cardiac-derived interstitial cell types from human heart tissue, as previously reported by our group, prompted design of a three-dimensional (3D) structure that maximizes cellular interaction, allows for defined cell ratios, controls size, enables injectability, and minimizes cell losses upon delivery. MethodsThree distinct populations of human cardiac interstitial cells including mesenchymal stem cells (MSCs), endothelial progenitor cells (EPCs), and c-Kit+ cardiac interstitial cells (cCICs) when cultured together spontaneously form scaffold-free 3D microenvironments termed CardioClusters. Biological consequences of CardioCluster formation were assessed by multiple assays including single cells RNA-Seq transcriptional profiling. Protective effects of CardioClusters in vitro were measured using cell culture models for oxidative stress and myocardial ischemia in combination with freshly isolated neonatal rat ventricular myocytes. Long-term impact of adoptively transferred CardioClusters upon myocardial structure and function in a xenogenic model of acute infarction using NODscid mice was assessed over a longitudinal time course of 20-weeks. ResultsCardioCluster design enables control over composite cell types, cell ratios, size, and preservation of structural integrity during delivery. Profound changes for biological properties of CardioClusters relative to constituent parental cell populations include enhanced expression of stem cell-relevant factors, adhesion/extracellular-matrix molecules, and cytokines. The CardioCluster 3D microenvironment maximizes cellular interaction while maintaining a more native transcriptome similar to endogenous cardiac cells. CardioCluster delivery improves cell retention following intramyocardial injection with preservation of long-term cardiac function relative to monolayer-cultured cells when tested in an experimental murine infarction model followed for up to 20 weeks post-challenge. CardioCluster-treated hearts show increases in capillary density, preservation of cardiomyocyte size, and reduced scar size indicative of blunting pathologic infarction injury. ConclusionsCardioClusters are a novel next generation development and delivery approach for cellular therapeutics that potentiate beneficial activity and enhance protective effects of human cardiac interstitial cell mixed populations. CardioClusters utilization in this preclinical setting establishes fundamental methodologic and biologic insights, laying the framework for optimization of CardioCluster design to provide greater efficacy in cell-based therapeutic interventions intended to mitigate cardiomyopathic damage.

cell biology↗

Tetraploidy in rodent cardiac stem cells confers enhanced biological properties

Ploidy for cardiomyocytes is well described but remains obscure in cardiac interstitial cells (CICs). Ploidy of c-kit+CICs were assessed using a combination of confocal, karyotypic, and flow cytometric assessments coupled with molecular and bioinformatic analyses. Fundamental differences were found between cultured rodent (rat, mouse) c-kit+CICs possessing mononuclear tetraploid (4n) content versus large mammal (human, swine) with mononuclear diploid (2n) content. In-situ analysis, confirmed with fresh isolates, revealed diploid content in c-kit+CICs from human and a mixture of diploid and tetraploid nuclei in mouse. Molecular assessment of the p53 signaling pathway provides a plausible explanation for escape from replicative senescence in rodent but not human ckit+CICs. Single cell transcriptional profiling reveals distinctions between diploid versus tetraploid populations in mouse ckit+CICs, alluding to functional divergences. Collectively, these data reveal fundamental species-specific biological differences in c-kit+CICs that could account for challenges in extrapolation of myocardial preclinical studies from rodent to large animal models.

cell biology↗

Safety profiling of genetically engineered Pim-1 kinase overexpression for oncogenicity risk in human c-kit+ cardiac interstitial cells

Advanced approaches to stem cell-based therapies is necessary for myocardial regenerative therapy because treatments have yielded modest results in the clinic. Our group previously demonstrated genetic modification of cardiac stem cells with Pim-1 kinase overexpression rejuvenated aged cells and potentiated myocardial repair. Despite these encouraging findings, concerns were raised regarding oncogenic risk associated with Pim-1 kinase overexpression. Testing of these c-kit+ cardiac interstitial cells (cCICs), derived from heart failure patient samples, overexpressing Pim-1 (cCICs-Pim-1) for indices of oncogenic risk was assessed by soft agar colony formation, micronucleation, gamma-Histone 2AX foci, and transcriptome profiling. Collectively, findings demonstrate comparable phenotypic and biological properties of cCICsPim-1 compared to baseline control cCICs with no evidence for oncogenic phenotype. Using a highly-selective and continuous sensor for quantitative assessment of PIM1 kinase activity, a 7-fold increase in cCICs-Pim-1 versus cCICs resulted. Kinase activity was elevated in IKKs, AKT/SGK, CDK1-3, p38, and ERK1/2 in addition to Pim-1, correlating Pim-1 overexpression to contribute to Pim-1-mediated effects. Enhancement of cellular survival, proliferation, and other beneficial properties to augment stem cell-mediated repair without oncogenic risk is a feasible, logical, and safe approach to improve efficacy and overcome current limitations inherent to cellular adoptive transfer therapeutic interventions.

cell biology↗