Cellular Adaptation Research Articles

XRT-06. RAPID PTEFB-DEPENDENT TRANSCRIPTIONAL REORGANIZATION UNDERPINS THE GLIOMA ADAPTIVE RESPONSE TO RADIOTHERAPY

Abstract Dynamic regulation of gene expression is fundamental for cellular adaptation to exogenous stressors. PTEFb-mediated pause-release of RNA polymerase II (Pol II) is a conserved regulatory mechanism for synchronous transcriptional induction in response to heat shock, but this pro-survival role has not been examined in the applied context of cancer therapy. Using a combination of ChIP-, ATAC-, and RNA-seq in model systems of diffuse intrinsic pontine glioma and other pediatric high-grade gliomas, we observe a rapid genome-wide reorganization of active chromatin within hours of exposure to therapeutic ionizing radiation. This redistribution facilitates PTEFb-mediated pause-release of Pol II to drive nascent transcriptional induction of canonical DNA damage response programs. Concurrent inhibition of PTEFb imparts a transcription elongation defect, disrupting this chromatin reorganization and blunting transcriptional induction. Functionally, concurrent PTEFb inhibition abrogates key adaptive programs such as DNA repair and cell cycle regulation. This combination demonstrates a potent, synergistic therapeutic potential agnostic of glioma subtype, leading to a marked induction of tumor cell apoptosis and prolongation of xenograft survival. These studies reveal a central role for PTEFb underpinning the early adaptive response to radiotherapy, opening new avenues for combinatorial treatment in these lethal malignancies.

Mitochondrial folate metabolism–mediated α-linolenic acid exhaustion masks liver fibrosis resolution

Sustainable TGF-β1 signaling drives organ fibrogenesis. However, the cellular adaptation to maintain TGF-β1 signaling remains unclear. In this study, we revealed that dietary folate restriction promoted the resolution of liver fibrosis in mice with nonalcoholic steatohepatitis. In activated hepatic stellate cells, folate shifted toward mitochondrial metabolism to sustain TGF-β1 signaling. Mechanistically, nontargeted metabolomics screening identified that α-linolenic acid (ALA) is exhausted by mitochondrial folate metabolism in activated hepatic stellate cells. Knocking down serine hydroxymethyltransferase 2 increases the bioconversion of ALA to docosahexaenoic acid, which inhibits TGF-β1 signaling. Finally, blocking mitochondrial folate metabolism promoted liver fibrosis resolution in nonalcoholic steatohepatitis mice. In conclusion, mitochondrial folate metabolism/ALA exhaustion/TGF-βR1 reproduction is a feedforward signaling to sustain profibrotic TGF-β1 signaling, and targeting mitochondrial folate metabolism is a promising strategy to enforce liver fibrosis resolution.

Effects of high-resistance wheel running on hallmarks of endurance and resistance training adaptations in mice.

Exercise effectively promotes and preserves cardiorespiratory, neuromuscular, metabolic, and cognitive functions throughout life. The molecular mechanisms underlying the beneficial adaptations to exercise training are, however, still poorly understood. To improve the mechanistic study of specific exercise training adaptations, standardized, physiological, and well-characterized training interventions are required. Therefore, we performed a comprehensive interrogation of systemic changes and muscle-specific cellular and molecular adaptations to voluntary low-resistance wheel running (Run) and progressive high-resistance wheel running (RR) in young male mice. Following 10 weeks of training, both groups showed similar improvements in body composition and peak oxygen uptake (V̇O2peak ), as well as elevated mitochondrial proteins and capillarization markers in the M. plantaris. Run mice clearly outperformed RR mice in a forced treadmill running capacity test, while RR mice displayed increased grip strength as well as superior mass gains in the M. soleus, associated with distinct proteomic changes specifying the two paradigms. Thus, even though both training modalities induce overlapping adaptations, Run interventions preferably improve submaximal running performance, while progressive RR is a valid model to study training-induced gains in grip strength and plantar flexor hypertrophy.

Loss of apolipoprotein E exacerbates microglial and astrocyte reactivity to sustained systemic glucocerebrosidase inhibition

AbstractBackgroundIncreased lipid storage is observed within the brains of patients with Alzheimer’s disease (AD). Genetic risk factors for AD involved in lipid processing include APOE, ABCA7 and TREM2. Lipid transport between neurons and glia is induced by mitochondrial stress and NMDA‐induced excitotoxicity in vitro and is dependent on glial apolipoprotein E (ApoE) expression. Therefore, this suggests that lipid transport between neurons and glia of the brain by ApoE and intracellular lipid processing are critical cellular adaptations to stress. This in turn suggests that insufficient ApoE activity may underpin the elevated lipid storage associated with neurodegenerative diseases.MethodTo determine whether ApoE function is required for cellular adaptation to increased lipid load relevant to dementia, 3‐month‐old WT and ApoE‐KO mice received daily intraperitoneal injections of the glucocerebrosidase inhibitor, CBE, for 18 d. Animals were sacrificed 1 d after the final dose and perfused with paraformaldehyde for immunohistochemistry, or were perfused with saline and dissected to isolate the cortex, hippocampus and substantia nigra for biochemical analysis by Western blot. Microglial and astrocyte reactivity to CBE were assessed by Iba1 and GFAP immunohistochemistry respectively.ResultMicroglial reactivity was greater in the dentate gyrus, substantia nigra and cortical layer V in ApoE‐KO mice after 18 d CBE, relative to WT. Loss of ApoE also elevated astrocyte reactivity in the dentate gyrus, substantia nigra and cortical layer V after 18 d CBE. Ongoing analyses into the molecular and cellular consequences of prolonged CBE include changes in lipid transport, tau phosphorylation, lysosome biogenesis, autophagy and proteostasis, to determine how loss of ApoE function may impact cellular adaptation to intracellular lipid formation resulting from sustained glucocerebrosidase inhibition.ConclusionThe exacerbated glial activation in ApoE‐KO animals receiving CBE suggests that ApoE expression and function may, in part, determine the capacity of neuron‐glia networks to overcome damaging stimuli. When ApoE function is insufficient, failed resolution of lipids may then lead to persistent damage and degeneration. Therefore, characterizing the consequences of limited lipid clearance on the adaptation to rising lipid load is highly relevant to understanding how failure to overcome increased lipid storage contributes to neurodegenerative processes in AD.

Energy restriction or improvements in diet quality: identifying the best pathway for a longer and healthier life.

Obesity is a major risk factor for chronic non-communicable diseases (NCDs) and it has increased to epidemic proportions. Unhealthy diet represents a modifiable risk factor for both obesity and NCDs, however, there is no universal dietary intervention to improve obesity-related NCDs and particularly to reduce the risk for major adverse cardiovascular events. Energy restriction (ER) and diet quality changes, with and without ER, have been widely investigated in preclinical and clinical studies, however, the potential underlying mechanisms driving the benefits of those dietary interventions remain largely unclear. ER affects multiple metabolic, physiological, genetic, and cellular adaptation pathways associated with prolonged lifespan, particularly in preclinical models, while these benefits remain to be established in humans. Moreover, the sustainability of ER and its implementation across the different diseases remains challenging. On the other hand, diet quality with improvements, with or without ER, has been associated with more favorable long-term metabolic and cardiovascular outcomes. This narrative review will describe the role of ER and/or diet quality improvements on the risk for NCDs. It will also discuss the potential mechanisms of action underlying the potential beneficial effects of those dietary approaches.

The mechanisms to dispose of misfolded proteins in the endoplasmic reticulum of adipocytes

Endoplasmic reticulum (ER)-associated degradation (ERAD) and ER-phagy are two principal degradative mechanisms for ER proteins and aggregates, respectively; however, the crosstalk between these two pathways under physiological settings remains unexplored. Using adipocytes as a model system, here we report that SEL1L-HRD1 protein complex of ERAD degrades misfolded ER proteins and limits ER-phagy and that, only when SEL1L-HRD1 ERAD is impaired, the ER becomes fragmented and cleared by ER-phagy. When both are compromised, ER fragments containing misfolded proteins spatially coalesce into a distinct architecture termed Coalescence of ER Fragments (CERFs), consisted of lipoprotein lipase (LPL, a key lipolytic enzyme and an endogenous SEL1L-HRD1 substrate) and certain ER chaperones. CERFs enlarge and become increasingly insoluble with age. Finally, we reconstitute the CERFs through LPL and BiP phase separation in vitro, a process influenced by both redox environment and C-terminal tryptophan loop of LPL. Hence, our findings demonstrate a sequence of events centered around SEL1L-HRD1 ERAD to dispose of misfolded proteins in the ER of adipocytes, highlighting the profound cellular adaptability to misfolded proteins in the ER in vivo.

Myosin and upalpha-actinin regulation of stress fiber contractility under tensile stress

Stress fibers are actomyosin bundles that regulate cellular mechanosensation and force transduction. Interacting with the extracellular matrix through focal adhesion complexes, stress fibers are highly dynamic structures regulated by myosin motors and crosslinking proteins. Under external mechanical stimuli such as tensile forces, the stress fiber remodels its architecture to adapt to external cues, displaying properties of viscoelastic materials. How the structural remodeling of stress fibers is related to the generation of contractile force is not well understood. In this work, we simulate mechanochemical dynamics and force generation of stress fibers using the molecular simulation platform MEDYAN. We model stress fiber as two connecting bipolar bundles attached at the ends to focal adhesion complexes. The simulated stress fibers generate contractile force that is regulated by myosin motors and alpha-actinin crosslinkers. We find that stress fibers enhance contractility by reducing the distance between actin filaments to increase crosslinker binding, and this structural remodeling ability depends on the crosslinker turnover rate. Under tensile pulling force, the stress fiber shows an instantaneous increase of the contractile forces followed by a slow relaxation into a new steady state. While the new steady state contractility after pulling depends only on the overlap between actin bundles, the short-term contractility enhancement is sensitive to the tensile pulling distance. We further show that this mechanical response is also sensitive to the crosslinker turnover rate. Our results provide new insights into the stress fiber mechanics that have significant implications for understanding cellular adaptation to mechanical signaling.

Remote support during cardiac implantable electrical device follow-up: preliminary experience in clinical practice

Abstract Funding Acknowledgements Type of funding sources: None. Background Industry employed allied professionals (IEAPs) traditionally provide technical assistance to physicians during cardiac implantable electrical device (CIED) implantation, programming, troubleshooting, and follow-up, by operating the programmers under the supervision of the physician. The Heart Connect™ application is a data-sharing system intended to provide physicians with the means to establish an online meeting and share the display of the LATITUDE™ Programmer (Boston Scientific) with IEAPs in a remote location. Purpose We describe the preliminary experience of remote IEAP support during CIED follow-up in clinical practice. Methods The Heart Connect™ software was downloaded on the programmer and network connection was established and tested using both Wi-Fi and a 4G cellular adapter at 3 Italian medical centers. External headset and webcam have been connected to the programmer. After system setup, the staff was trained on how to use the application, and online meetings were established with IEAPs of the Remote Clinical Support team during consecutive CIED follow-up visits. Data and users’ feedback were collected on the training provided, the quality of the remote connection, the support at each meeting. Results All the professionals trained in the use of the system judged the information received to be clear and adequate, and felt confident in using the system. Online meetings were established during 34 follow-up visits (12 pacemakers and CRT-P, 13 single- or dual-chamber ICD, 9 CRT-D). The remote connection was successfully established in all cases. In all cases the communication remained stable for the entire duration of the visit, except for 2 cases (connected via 4G) in which a loss of connection occurred (not restored in 1 case). The sound and video quality during two-way communication were rated good or excellent in 31 (91%) cases. The mean duration of the online meeting was 7±3 minutes. Routine CIED checks were performed during the visits: battery status, pacing and sensing thresholds, impedances, current rhythm diagnosis, arrhythmias detected and therapies delivered, device counters and review of programmed parameters, alerts and clinical diagnostics. Programming changes were done during 7 (21%) visits: pacing and sensing parameters, tachycardia detection and therapies. The remote support allowed to successfully complete the visit in 33 (97%) cases, without requiring additional medical or technical support. In all these cases, the support was judged to be equally effective compared to the on-site support, without causing delays or organizational difficulties. Conclusions Remote support during CIED follow-up using Heart Connect™ seems feasible, effective and well accepted. Additional tests are ongoing to demonstrate the potential usefulness of this solution in standard clinical practice, and in the context of restrictions on access to healthcare facilities due to COVID-19 pandemic.

Evolution, mechanism and limits of dietary restriction induced health benefits & longevity

Dietary restriction (DR) is the most powerful intervention to enhance health and lifespan across species. However, recent findings indicate that DR started in late life has limited capacity to induce health benefits. Age-dependent changes that impair DR at old age remain to be delineated. This requires a better mechanistic understanding of the different aspects that constitute DR, how they act independently and in concert. Current research efforts aim to tackle these questions: Are fasting periods needed for the induction of DR's health benefits? Does the improvement of cellular and organismal functions depend on the reduction of specific dietary components like proteins or even micronutrients and/or vitamins? How is the aging process intervening with DR-mediated responses? Understanding the evolutionary benefits of nutrient stress responses in driving molecular and cellular adaptation in response to nutrient deprivation is likely providing answers to some of these questions. Cellular memory of early life may lead to post-reproductive distortions of gene regulatory networks and metabolic pathways that inhibit DR-induced stress responses and health benefits when the intervention is started at old age. Inhere we discuss new insights into mechanisms of DR-mediated health benefits and how evolutionary selection for fitness in early life may limit DR-mediated improvements at old age.

A long-acting GDF15 analog causes robust, sustained weight loss and reduction of food intake in an obese nonhuman primate model.

Growth Differentiation Factor-15 (GDF15) is a circulating polypeptide linked to cellular stress and metabolic adaptation. GDF15's half-life is ~3 h and activates the glial cell line-derived neurotrophic factor family receptor alpha-like (GFRAL) receptor expressed in the area postrema. To characterize sustained GFRAL agonism on food intake (FI) and body weight (BW), we tested a half-life extended analog of GDF15 (Compound H [CpdH]) suitable for reduced dosing frequency in obese cynomolgus monkeys. Animals were chronically treated once weekly (q.w.) with CpdH or long-acting GLP-1 analog dulaglutide. Mechanism-based longitudinal exposure-response modeling characterized effects of CpdH and dulaglutide on FI and BW. The novel model accounts for both acute, exposure-dependent effects reducing FI and compensatory changes in energy expenditure (EE) and FI occurring over time with weight loss. CpdH had linear, dose-proportional pharmacokinetics (terminal half-life ~8 days) and treatment caused exposure-dependent reductions in FI and BW. The 1.6 mg/kg CpdH reduced mean FI by 57.5% at 1 week and sustained FI reductions of 31.5% from weeks 9-12, resulting in peak reduction in BW of 16 ± 5%. Dulaglutide had more modest effects on FI and peak BW loss was 3.8 ± 4.0%. Longitudinal modeling of both the FI and BW profiles suggested reductions in BW observed with both CpdH and dulaglutide were fully explained by exposure-dependent reductions in FI without increase in EE. Upon verification of the pharmacokinetic/pharmacodynamic relationship established in monkeys and humans for dulaglutide, we predicted that CpdH could reach double digit BW loss in humans. In summary, a long-acting GDF15 analog led to sustained reductions in FI in overweight monkeys and holds potential for effective clinical obesity pharmacotherapy.

VBP1 negatively regulates CHIP and selectively inhibits the activity of hypoxia-inducible factor (HIF)-1α but not HIF-2α

Hypoxia-inducible factor-1 (HIF-1) is a critical transcription factor that regulates the expression of genes involved in cellular adaptation to low oxygen levels. Aberrant regulation of the HIF-1 signaling pathway is linked to various human diseases. Previous studies have established that HIF-1α is rapidly degraded in a von Hippel-Lindau protein (pVHL)-dependent manner under normoxic conditions. In this study, we find that pVHL binding protein 1 (VBP1) is a negative regulator of HIF-1α but not HIF-2α using zebrafish as an invivo model and invitro cell culture models. Deletion of vbp1 in zebrafish caused Hif-1α accumulation and upregulation of Hif target genes. Moreover, vbp1 was involved in the induction of hematopoietic stem cells (HSCs) under hypoxic conditions. However, VBP1 interacted with and promoted the degradation of HIF-1α in a pVHL-independent manner. Mechanistically, we identify the ubiquitin ligase CHIP and HSP70 as new VBP1 binding partners and demonstrate that VBP1 negatively regulated CHIP and facilitated CHIP-mediated degradation of HIF-1α. In patients with clear cell renal cell carcinoma (ccRCC), lower VBP1 expression was associated with worse survival outcomes. In conclusion, our results link VBP1 with CHIP stability and provide insights into underlying molecular mechanisms of HIF-1α-driven pathological processes.

Activation of the PERK branch of the unfolded protein response during production reduces specific productivity in CHO cells via downregulation of PDGFRa and IRE1a signaling.

During the course of biopharmaceutical production, heterologous protein expression in Chinese hamster ovary (CHO) cells imposes a high proteostatic burden that requires cellular adaptation. To mitigate such burden, cells utilize the unfolded protein response (UPR), which increases endoplasmic reticulum (ER) capacity to accommodate elevated rates of protein synthesis and folding. In this study, we show that during production the UPR regulates growth factor signaling to modulate growth and protein synthesis. Specifically, the protein kinase R-like ER kinase (PERK) branch of the UPR is responsible for transcriptional down-regulation of platelet-derived growth factor receptor alpha (PDGFRa) and attenuation of the IRE1-alpha (IRE1a) branch of the UPR. PERK knockout (KO) cell lines displayed reduced growth and viability due to higher rates of apoptosis despite having stabilized PDGFRa levels. Knocking out PERK in an apoptosis impaired (Bax/Bak double KO) antibody-expressing cell line prevented apoptotic cell death and revealed that apoptosis was likely triggered by increased ER stress and reactive oxygen species levels in the PERK KO hosts. Our findings suggest that attenuation of IRE1a and PDGFRa signaling by the PERK branch of the UPR reduces ER protein folding capacity and hence specific productivity of CHO cells in order to mitigate UPR and prevent apoptotic cell death. Last, Bax/Bak/PERK triple KO CHO cell lines displayed 2-3 folds higher specific productivity and titer (up to 8 g/L), suggesting that modulation of PERK signaling during production processes can greatly improve specific productivity in CHO cells.

α-Ketoglutarate stimulates cell growth through the improvement of glucose and glutamine metabolism in C2C12 cell culture.

Cellular adaptation to physical training and energy metabolism play an important role during physical exercise. This study sought to investigate the effects of α-KG on cell growth and energy metabolism in C2C12 cell culture. C2C12 cells were cultured in media pretreated without (control) or with α-KG at different concentrations, and cells and media were harvested every 24 h for 8 days. From cell counts, specific cell growth rate (SGR) and doubling time were calculated. The content of glucose, glutamine, lactate, and ammonia in media was determined, and the specific consumption rate (SCR) or production rate (SPR) was calculated. Additionally, cell colony-forming efficiency (CFE) was determined. The control cells showed a CFE at 50%, a typical cell growth curve in the first 5 days with a mean SGR at 0.86/day, and a mean cell count doubling time at 19.4 h. In the group with α-KG at 100 mM, the cells underwent rapid cell death, and thus no further analysis was made. The treatment with α-KG at lower concentrations (0.1 mM and 1.0 mM) led to a higher CFE at 68 and 55%, respectively, whereas those in groups with higher α-KG concentration decreased (10 and 6% for 20 mM and 30 mM α-KG, respectively). The mean SGR was 0.95/day, 0.94/day, 0.77/day, 0.71/day, and 0.65/day for groups treated with α-KG at 0.1, 1.0, 10.0, 20.0, and 30.0 mM, respectively, and the corresponding cell count doubling time was 17.6, 17.8, 20.9, 24.6, and 24.7 h, respectively. In comparison with that of the control group, the mean glucose SCR decreased in all the groups treated with α-KG, while the mean glutamine SCR remained unchanged; the mean lactate SPR increased in the groups treated with α-KG ≥ 20.0 mM. Finally, the mean SPR of ammonia was lower in all α-KG groups than that in the control. The treatment with α-KG at lower concentrations increased cell growth whereas at higher concentrations decreased cell growth, and α-KG reduced glucose consumption and ammonia production. Therefore, α-KG stimulates cell growth in a dose-dependent manner, which is likely through the improvement of glucose and glutamine metabolism in a C2C12 culture setting.

BRAF V600-Mutated Metastatic Melanoma and Targeted Therapy Resistance: An Update of the Current Knowledge.

Melanoma is the most common cause of death in skin cancer due to its high metastatic potential. While targeted therapies have improved the care of patients with metastatic melanoma harboring the BRAFV600E mutation, these treatments are associated with a high frequency of resistance. Resistance factors are related to cellular adaptation as well as to changes in the tumor microenvironment. At the cellular level, resistance involves mutations, overexpression, activation, or inhibition of effectors involved in cell signaling pathways such as MAPK, PI3K/AKT, MITF, and epigenetic factors (miRNAs). In addition, several components of the melanoma microenvironment, such as soluble factors, collagen, and stromal cells also play a crucial role in this resistance. In fact, extracellular matrix remodeling impacts the physical and chemical properties with changes in the stiffness and acidity, respectively of the microenvironment. The cellular and immune components of the stroma are also affected, including immune cells and CAF. The aim of this manuscript is to review the mechanisms responsible for resistance to targeted therapies in BRAFV600E-mutated metastatic melanoma.

Mechanisms of Cellular Adaptation and Change: A Narrative Literature Review

Injury to cells and the surrounding environment, called the extracellular matrix, triggers injury to tissues and organs. Although a normal cell is limited by narrow boundaries of structure and function, it is capable of adapting to biological demands or stress to maintain a steady state called homeostasis. Adaptation is a reversible, structural, or functional response to normal or physiological conditions and adverse or pathological conditions. This review aimed to describe the mechanism of cellular adaptation in the human body. Cells adapt to the environment to escape and protect against injury. Adaptation of the cell, be it normal or injured, this condition lies somewhere between these two conditions. The most significant adaptive changes in cells include atrophy (decreased cell size), hypertrophy (increased cell size), hyperplasia (increased cell number), and metaplasia (reversible replacement of one mature cell for another less mature cell or change in phenotype). Dysplasia (a disorder of cellular growth) is not considered a true cellular adaptation but rather an atypical hyperplasia. In conclusion, cellular adaptation is a central and common part of many disease conditions.

Stress granules: potential therapeutic targets for infectious and inflammatory diseases.

Eukaryotic cells are stimulated by external pressure such as that derived from heat shock, oxidative stress, nutrient deficiencies, or infections, which induce the formation of stress granules (SGs) that facilitates cellular adaptation to environmental pressures. As aggregated products of the translation initiation complex in the cytoplasm, SGs play important roles in cell gene expression and homeostasis. Infection induces SGs formation. Specifically, a pathogen that invades a host cell leverages the host cell translation machinery to complete the pathogen life cycle. In response, the host cell suspends translation, which leads to SGs formation, to resist pathogen invasion. This article reviews the production and function of SGs, the interaction between SGs and pathogens, and the relationship between SGs and pathogen-induced innate immunity to provide directions for further research into anti-infection and anti-inflammatory disease strategies.

Abstract #1404234: Hypoglycemia & Lactic Acidosis Due to Warburg Effect

Warburg effect (Aerobic Glycolysis) is a form of cellular adaptation by cancer cells to support its biosynthetic requirement, when in the presence of abundant oxygen and mitochondria, cell utilizes anaerobic pathway to convert glucose to lactate producing only 2 ATP’s. This phenomenon has been studied extensively and clinically can results in high glucose uptake by the tumor cells causing hypoglycemia and type B lactic acidosis. 41-year-old male with no PMHx admitted with weakness, instability, vision changes and unintentional 30 lbs weight loss x 3 months. In the ER he was found to be have BP 90/60, HR 113 and afebrile with Glucose of 20, lab showed AKI with lactic acidosis (Cr. 2.17 and lactic acid >10) and CT Abd/Plv showed multiple liver metastatic disease and entrocolitis. Started on IV fluids with dextrose, Pressor support, Antibiotics and admitted to ICU for care. Workup showed HIV positive and antibiotics coverage was broadened. Kidney function continued to worsen and started on CRRT. Patient’s hypoglycemia persisted despite dextrose infusion and endocrine team was consulted. Work up showed appropriately low insulin & c-peptide with negative sulfonylurea screen. Biopsy of liver confirmed Large B-Cell Lymphoma, but not a candidate for therapy due to acute illness. Patient lactic acidosis persisted >10 despite therapy suggestive of Type B lactic acidosis. Family changed goals of care & patient passed away within 48 hours of comfort care measures. Presence of hypoglycemia and lactic acidosis is a medical emergency suggesting hyper metabolic state with poor tissue oxygenation, mostly caused by severe inflammatory resulting in Type A Lactic Acidosis. Rarely this is seen in normoxic condition called Type B Lactic acidosis and one of the potential causes is Warburg effects, in patients with solid and hematological malignancy. This is triggered by oncogenic lesions such as expression of hypoxia-inducible factor-1α (HIF1α) causing up regulation of major glycolytic enzymes like hexokinase and Tumor M2 Pyruvate Kinase. This provides cancer cell with metabolic intermediates required for synthesis of cellular components for cell proliferation and also produces less amounts of free radicals. Proliferating cancer cells also outpace the vascular supply and warburg effect ensures survival in the hypoxic environment. Patients with type B lactic acidosis and hypoglycemia poses grim prognosis with mortality around 75% (7 out of 29 patient survivals, in chart review of cases). Therapeutic measure includes: tumor directed therapies, IV Bicarb Infusion, IV Dextrose, Renal Replacement Therapy and Thiamine replacement. Purpose of this case presentation is to further enhance clinical evidence of Warburg effect.

The HIF-prolyl hydroxylases have distinct and non-redundant biological roles in the pathogenesis of colitis-associated cancer

Colitis-associated colorectal cancer (CAC) is a severe complication of inflammatory bowel disease (IBD). HIF-prolyl hydroxylases (PHD1, PHD2, and PHD3) control cellular adaption to hypoxia and are considered promising therapeutic targets in IBD. However, their relevance in the pathogenesis of CAC remains elusive. We induced CAC in Phd1-/-, Phd2+/-, Phd3-/-, and WT mice with azoxymethane (AOM) and dextran sodium sulfate (DSS). Phd1-/- mice were protected against chronic colitis and displayed diminished CAC growth compared to WT mice. In Phd3 -/- mice, colitis activity and CAC growth remained unaltered. In Phd2+/- mice, colitis activity was unaffected, but CAC growth was aggravated. Mechanistically, Phd2 deficiency (i) increased the number of tumor-associated macrophages (TAMs) in AOM/DSS-induced tumors, (ii) promoted the expression of EGFR ligand epiregulin ( Ereg) in macrophages, and (iii) augmented signal transducer and activator of transcription 3 (STAT3) and extracellular signal-regulated kinase 1/2 (ERK1/2) signaling which at least in part contributed to aggravated tumor cell proliferation in colitis-associated tumors. Consistently, Phd2 deficiency in hematopoietic ( Vav:Cre-Phd2f/f) but not in intestinal epithelial cells ( Villin:Cre-Phd2f/f) increased CAC growth. In conclusion, the three different PHD isoenzymes have distinct and non-redundant, either promoting (PHD1), diminishing (PHD2), or neutral effects (PHD3) on CAC growth. KBK received funding from the German Cancer Aid. MJS. received funding from the German Research Foundation (DFG; STR 1570/1-1) and the Braun Foundation (Braun®; BBST-D-18-00018). MSch received funding from the German Federal Ministry of Education and Research (BMBF; 031L0084). JMH received funding from the Heidelberg Stiftung Chirurgie. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

CHRNA5 gene variation affects the response of VTA dopaminergic neurons during chronic nicotine exposure and withdrawal

Nicotine is the principal psychoactive component in tobacco that drives addiction through its action on neuronal nicotinic acetylcholine receptors (nAChR). The nicotinic receptor gene CHRNA5, which encodes the α5 subunit, is associated with nicotine use and dependence. In humans, the CHRNA5 missense variant rs16969968 (G > A) is associated with increased risk for nicotine dependence and other smoking-related phenotypes. In rodents, α5-containing nAChRs in dopamine (DA) neurons within the ventral tegmental area (VTA) powerfully modulate nicotine reward and reinforcement. Although the neuroadaptations caused by long-term nicotine exposure are being actively delineated at both the synaptic and behavioral levels, the contribution of α5-containing nAChRs to the cellular adaptations associated with long-term nicotine exposure remain largely unknown. To gain insight into the mechanisms behind the influence of α5-containing nAChRs and the rs16969968 polymorphism on nicotine use and dependence, we used electrophysiological approaches to examine changes in nAChR function arising in VTA neurons during chronic nicotine exposure and multiple stages of nicotine withdrawal. Our results demonstrate that CHRNA5 mutation leads to profound changes in VTA nAChR function at baseline, during chronic nicotine exposure, and during short-term and prolonged withdrawal. Whereas nAChR function was suppressed in DA neurons from WT mice undergoing withdrawal relative to drug-naïve or nicotine-drinking mice, α5-null mice exhibited an increase in nAChR function during nicotine exposure that persisted throughout 5–10 weeks of withdrawal. Re-expressing the hypofunctional rs16969968 CHRNA5 variant in α5-null VTA DA neurons did not rescue the phenotype, with α5-SNP neurons displaying a similar increased response to ACh during nicotine exposure and early stages of withdrawal. These results demonstrate the importance of VTA α5-nAChRs in the response to nicotine and implicate them in the time course of withdrawal.

Mesenchymal Stem Cell Behavior under Microgravity: From Stress Response to a Premature Senescence.

Mesenchymal stem cells are undifferentiated cells able to acquire different phenotypes under specific stimuli. Wharton's jelly is a tissue in the umbilical cord that contains mesenchymal stromal cells (MSCs) with a high plasticity and differentiation potential. Their regeneration capability is compromised by cell damage and aging. The main cause of cell damage is oxidative stress coming from an imbalance between oxidant and antioxidant species. Microgravity represents a stressing condition able to induce ROS production, ultimately leading to different subcellular compartment damages. Here, we analyzed molecular programs of stemness (Oct-4; SOX2; Nanog), cell senescence, p19, p21 (WAF1/CIP1), p53, and stress response in WJ-MSCs exposed to microgravity. From our results, we can infer that a simulated microgravity environment is able to influence WJ-MSC behavior by modulating the expression of stress and stemness-related genes, cell proliferation regulators, and both proapoptotic and antiapoptotic genes. Our results suggest a cellular adaptation addressed to survival occurring during the first hours of simulated microgravity, followed by a loss of stemness and proliferation capability, probably related to the appearance of a molecular program of senescence.