Biological Hallmarks Research Articles

SARCOPENIA IN OLDER ADULTS UNDERGOING CARDIAC SURGERY

Sarcopenia, defined as age-related loss of muscle mass and strength, is one of the biological hallmarks of frailty. In practice, muscle mass is typically measured using a dual x-ray absorptiometry (DXA) scan, while strength is measured using physical performance tests. However, DXA may be less accurate in acute cardiac patients due to the confounding effect of body edema, and strength alone may be a superior indicator of muscle quality in aging. A convenience sample of older adults referred for cardiac surgery were prospectively consented and enrolled at the Jewish General Hospital. After a structured questionnaire and physical performance battery, patients underwent a DXA scan (GE Lunar) to measure their appendicular muscle mass (AMM) – the sum of fat-free tissue in their arms and legs. Patients were categorized as sarcopenic based on the European Working Group guidelines if they had low AMM and low strength as measured by the timed chair rise test >15 seconds. Both non-indexed and height-squared-indexed cutoffs were explored for low AMM. A Cox proportional hazards model was used to test the association between sarcopenia (or its individual components) and all-cause mortality adjusting for age, sex, and cardiac surgery type. The cohort consisted of 134 patients with a mean age of 70.7 ± 10.2 years and 23% female. The cardiac surgery type was isolated coronary bypass in 58%, valve surgery in 30%, and decision not to proceed with surgery in 12%. The mean AMM was 24.2 ± 5.1 kg in men and 21.0 ± 5.2 kg in women. The prevalence of sarcopenia was 10% and 13% using non-indexed and indexed cutoffs, respectively, similar in men and women. In the multivariable model, sarcopenia was not associated with mortality (N=24) over 2 years of follow-up, and neither was AMM alone. Timed chair rise time alone was associated with mortality as a continuous variable and a dichotomous >15 second variable (HR 7.72, 95% CI 1.75 to 34.11). Lower-extremity muscle strength is more predictive than muscle mass or DXA-based sarcopenia in predicting survival after cardiac surgery.

Candidate Genes and MiRNAs Linked to the Inverse Relationship Between Cancer and Alzheimer's Disease: Insights From Data Mining and Enrichment Analysis.

The incidence of cancer and Alzheimer’s disease (AD) increases exponentially with age. A growing body of epidemiological evidence and molecular investigations inspired the hypothesis of an inverse relationship between these two pathologies. It has been proposed that the two diseases might utilize the same proteins and pathways that are, however, modulated differently and sometimes in opposite directions. Investigation of the common processes underlying these diseases may enhance the understanding of their pathogenesis and may also guide novel therapeutic strategies. Starting from a text-mining approach, our in silico study integrated the dispersed biological evidence by combining data mining, gene set enrichment, and protein-protein interaction (PPI) analyses while searching for common biological hallmarks linked to AD and cancer. We retrieved 138 genes (ALZCAN gene set), computed a significant number of enriched gene ontology clusters, and identified four PPI modules. The investigation confirmed the relevance of autophagy, ubiquitin proteasome system, and cell death as common biological hallmarks shared by cancer and AD. Then, from a closer investigation of the PPI modules and of the miRNAs enrichment data, several genes (SQSTM1, UCHL1, STUB1, BECN1, CDKN2A, TP53, EGFR, GSK3B, and HSPA9) and miRNAs (miR-146a-5p, MiR-34a-5p, miR-21-5p, miR-9-5p, and miR-16-5p) emerged as promising candidates. The integrative approach uncovered novel miRNA-gene networks (e.g., miR-146 and miR-34 regulating p62 and Beclin1 in autophagy) that might give new insights into the complex regulatory mechanisms of gene expression in AD and cancer.

Ageing as a risk factor for neurodegenerative disease.

Ageing is the primary risk factor for most neurodegenerative diseases, including Alzheimer disease (AD) and Parkinson disease (PD). One in ten individuals aged ≥65 years has AD and its prevalence continues to increase with increasing age. Few or no effective treatments are available for ageing-related neurodegenerative diseases, which tend to progress in an irreversible manner and are associated with large socioeconomic and personal costs. This Review discusses the pathogenesis of AD, PD and other neurodegenerative diseases, and describes their associations with the nine biological hallmarks of ageing: genomic instability, telomere attrition, epigenetic alterations, loss of proteostasis, mitochondrial dysfunction, cellular senescence, deregulated nutrient sensing, stem cell exhaustion and altered intercellular communication. The central biological mechanisms of ageing and their potential as targets of novel therapies for neurodegenerative diseases are also discussed, with potential therapies including NAD+ precursors, mitophagy inducers and inhibitors of cellular senescence.

Evaluation of SOX2 and podoplanin expression in oral epithelial dysplasia and its correlation with malignant transformation.

Oral carcinogenesis cascade is a complex process, characterized by variable numbers of genetic and epigenetic alterations of various genes with manifold roles that could serve as biological hallmarks. This study was undertaken to assess the protein expression of SOX2 and podoplanin in oral epithelial dysplasia and correlate the expression with clinicopathological parameters and risk of malignant transformation. SOX2 and podoplanin expression were analyzed in 60 cases of oral epithelial dysplasia. The association between SOX2 and podoplanin expression with various clinicopathological parameters and transformation to oral cancer was analyzed. A higher Histoscore was seen in 55% of moderate and 30% of severe dysplasia. 25% of the cases showed a negative podoplanin expression and 30% of patients had higher podoplanin expression (score 2 and 3). Though there was significant association of both SOX2 and podoplanin expression with the degree of dysplasia, the association of their expression with transformation to oral squamous cell carcinoma did not reach statistical significance. Alteration in SOX2 and podoplanin is likely an important event in head and neck carcinogenesis; however, their expression may be valuable only in a few cases of oral epithelial dysplasia to assess the risk of malignant transformation.

A Clinically Applicable Gene-Expression Classifier Reveals Intrinsic and Extrinsic Contributions to Consensus Molecular Subtypes in Primary and Metastatic Colon Cancer.

Four consensus molecular subtypes (CMS1-4) of colorectal cancer were identified in primary tumors and found to be associated with distinctive biological features and clinical outcomes. Given that distant metastasis largely accounts for colorectal cancer-related mortality, we examined the molecular and clinical attributes of CMS in metastatic colorectal cancer (mCRC). We developed a colorectal cancer-focused NanoString-based CMS classifier that is ideally suited to interrogate archival tissues. We successfully used this panel in the CMS classification of formalin-fixed paraffin-embedded (FFPE) tissues from mCRC cohorts, one of which is composed of paired primary tumors and metastases. Finally, we developed novel mouse implantation models to enable modeling of colorectal cancer in vivo at relevant sites. Using our classifier, we find that the biological hallmarks of mCRC, including CMS, are in general highly similar to those observed in nonmetastatic early-stage disease. Importantly, our data demonstrate that CMS1 has the worst outcome in relapsed disease, compared with other CMS. Assigning CMS to primary tumors and their matched metastases reveals mostly concordant subtypes between primary and metastasis. Molecular analysis of matched discordant pairs reveals differences in stromal composition at each site. The development of two novel in vivo orthotopic implantation models further reinforces the notion that extrinsic factors may impact on CMS identification in matched primary and metastatic colorectal cancer. We describe the utility of a NanoString panel for CMS classification of FFPE clinical samples. Our work reveals the impact of intrinsic and extrinsic factors on colorectal cancer heterogeneity during disease progression.

Single-Cell Signature Explorer for comprehensive visualization of single cell signatures across scRNA-seq datasets.

The momentum of scRNA-seq datasets prompts for simple and powerful tools exploring their meaningful signatures. Here we present Single-Cell_Signature_Explorer (https://sites.google.com/site/fredsoftwares/products/single-cell-signature-explorer), the first method for qualitative and high-throughput scoring of any gene set-based signature at the single cell level and its visualization using t-SNE or UMAP. By scanning datasets for single or combined signatures, it rapidly maps any multi-gene feature, exemplified here with signatures of cell lineages, biological hallmarks and metabolic pathways in large scRNAseq datasets of human PBMC, melanoma, lung cancer and adult testis.

Abstract 3527: Combined high-throughput DNA genotyping and protein quantification in single cancer cells

Abstract Clonal genetic diversity is a hallmark of tumor biology. In many cancers, clonal evolution has been implicated as a driver of tumor initiation, progression, and relapse, and can be a prognostic indicator of disease outcome. Cancer cells are also phenotypically heterogeneous, constituting features which can enable intravasation and metastasis, or which can impart immunoevasive capabilities. Clinically, a combination of single-cell genotype and phenotype data from cancer biopsies could provide valuable insights into oncological disease states, such as metastatic risk or drug response. Here we present a multiomic technology for the simultaneous, high-throughput capture of DNA genotype and protein information from single cells, demonstrated through the analysis of acute myeloid leukemia (AML) derived cell lines. The experimental workflow leverages the high-throughput single-cell microfluidic droplet technology on the Mission Bio Tapestri platform, a system optimized to conduct multiplex targeted genomic DNA amplification from >50 AML-associated loci. In our modified workflow, cells are stained with a pool of monoclonal antibodies conjugated with barcoded oligonucleotides, enabling a sequencing-based readout of single-cell protein signatures. The sequencing data is analyzed with a bioinformatic pipeline that separates antibody signal from targeted genotyping data on a cell-by-cell basis. We find that on a mixed population of myelogenous leukemia cells and B-cells, single-cell genotype information effectively distinguishes between cell lines, while the protein data independently recapitulates the biological signal from conventional flow cytometric methods. In conclusion, this technology opens up a new era for discovery using multiomic analysis of the complex relationships between genotype and phenotype in cancer, paving the way for future studies analyzing primary tissue samples. Citation Format: Benjamin Demaree, Cyrille Delley, David Ruff, Sebastian Treusch, Dennis Eastburn, Keith Jones, Adam Abate. Combined high-throughput DNA genotyping and protein quantification in single cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3527.

Frontiers in positron emission tomography imaging of the vulnerable atherosclerotic plaque.

Rupture of vulnerable atherosclerotic plaques leading to an atherothrombotic event is the primary driver of myocardial infarction and stroke. The ability to detect non-invasively the presence and evolution of vulnerable plaques could have a huge impact on the future identification and management of atherosclerotic cardiovascular disease. Positron emission tomography (PET) imaging with an appropriate radiotracer has the potential to achieve this goal. This review will discuss the biological hallmarks of plaque vulnerability before going on to evaluate and to present PET imaging approaches which target these processes. The focus of this review will be on techniques beyond [18F]FDG imaging, some of which are clinically advanced, and others which are on the horizon. As inflammation is the primary driving force behind atherosclerotic plaque development, we will predominantly focus on approaches which either directly, or indirectly, target this process.

PS1447 ANALYSIS OF UPR‐RELATED GENE EXPRESSION IN CD34+ CELLS FROM PATIENTS WITH PRIMARY MYELOFIBROSIS REVEALS DOWNREGULATION OF UPR GENES AND REDUCTION OF APOPTOSIS

Background:Primary myelofibrosis (PMF) is a Philadelphia‐negative chronic myeloproliferative disorder characterized by splenomegaly, bone marrow fibrosis and a variable degree of myeloproliferation that can affect one or more hematopoietic lineages. The activation of the JAK/STAT pathway of the hematopoietic cells is a biological hallmark of the disease and is associated to acquired mutations that occur in JAK2, CALR or MPL genes. JAK/STAT pathway activation gives reason of the myeloproliferation; however, it does not completely explain how and why the pathological clone(s) emerge and expand in the bone marrow of PMF patients. The unfolded protein response (UPR) is an endoplasmic reticulum (ER)‐ specific pathway comprising a network of proteins that are activated in response to ER stress, due to accumulation of misfolded proteins. This can result from the exposure of the cell to stress stimuli, such as ROS overproduction, or DNA damage. UPR activation can result either in the recovery of the “stressed” cell through protein synthesis reduction and clearing of misfolded proteins or, if homeostasis is not restored, in its elimination through the activation of the apoptotic cascade. Recently, it has been reported that UPR activation maintains the integrity of hematopoietic stem/progenitor cells of healthy subjects by a balance between cell repair and induction of apoptosis (van Galen, P. et al.Nature 2014)Aims:To assess the activation of the UPR pathway in CD34+ hematopoietic progenitor cells obtained from patients with PMF, by investigating the expression of the main components of the UPR, including those implicated in the apoptotic cascade.Methods:RNA was obtained from CD34+ cells from the peripheral blood of 8 PMF patients (3 with a JAK2 p.V617F mutation, 2 with “Type 1” and 3 with “Type 2” CALR mutation) and 4 healthy subjects. Reverse transcription droplet digital PCR (RT‐ddPCR) was used to analyze the expression of the main genes belonging to the three branches of the UPR: ATF4, DDIT3, DNAJB9 (as representative of PERK branch), XBP1 s, XBP1u, HSPA5 (IRE1‐alpha branch) and ATF6 (ATF6 branch). The expression of BAX and BCL2 (involved in apoptosis) and of WT1 (as a measure of the myeloproliferative activity) was also analyzed.Results:As reported in Figure 1a, a significant downregulation of the genes belonging to the PERK and IRE1‐alpha branches of the UPR pathway was observed in CD34+ cells from PMF patients compared to healthy subjects. In particular, CD34+ cells from CALR‐mutated patients shows the lowest levels of expression in addition to downregulation of ATF6 (Figure 1b), whereas in JAK2V617F‐mutated patients, the ratio between the spliced and unspliced form of XBP1, that measures the activity of the IRE1‐alpha branch of UPR, is lower than in healthy subjects. This is accompanied by a significant upregulation of WT1 (Figure 1c). In CD34+ cell samples from all patients was also observed a reduced BAX/BCL2 ratio, mainly due to an increase of BCL2 expression.Summary/Conclusion:Since UPR‐dependent apoptosis activation can be induced both by ATF4‐mediated BCL2 downregulation and by IRE1alpha‐mediated BAX up‐regulation, our data strongly suggest that UPR‐mediated apoptosis is suppressed in hematopoietic progenitor cells from PMF patients. In keeping with this observation, the expression of WT1, a pro‐proliferative gene, is increased in the same cells. We hypothesize that UPR downregulation, by abolishing a pre‐differentiation checkpoint at stem/progenitor cell level, favors the emergence of the pathological clones.image

Regional perturbation of gene transcription is associated with intrachromosomal rearrangements and gene fusion transcripts in high grade ovarian cancer

Genomic rearrangements are a hallmark of cancer biology and progression, allowing cells to rapidly transform through alterations in regulatory structures, changes in expression patterns, reprogramming of signaling pathways, and creation of novel transcripts via gene fusion events. Though functional gene fusions encoding oncogenic proteins are the most dramatic outcomes of genomic rearrangements, we investigated the relationship between rearrangements evidenced by fusion transcripts and local expression changes in cancer using transcriptome data alone. 9,953 gene fusion predictions from 418 primary serious ovarian cancer tumors were analyzed, identifying depletions of gene fusion breakpoints within coding regions of fused genes as well as an N-terminal enrichment of breakpoints within fused genes. We identified 48 genes with significant fusion-associated upregulation and furthermore demonstrate that significant regional overexpression of intact genes in patient transcriptomes occurs within 1 megabase of 78 novel gene fusions that function as central markers of these regions. We reveal that cancer transcriptomes select for gene fusions that preserve protein and protein domain coding potential. The association of gene fusion transcripts with neighboring gene overexpression supports rearrangements as mechanism through which cancer cells remodel their transcriptomes and identifies a new way to utilize gene fusions as indicators of regional expression changes in diseased cells with only transcriptomic data.

Long non-coding RNAs as pan-cancer master gene regulators of associated protein-coding genes: a systems biology approach.

Despite years of research, we are still unraveling crucial stages of gene expression regulation in cancer. On the basis of major biological hallmarks, we hypothesized that there must be a uniform gene expression pattern and regulation across cancer types. Among non-coding genes, long non-coding RNAs (lncRNAs) are emerging as key gene regulators playing powerful roles in cancer. Using TCGA RNAseq data, we analyzed coding (mRNA) and non-coding (lncRNA) gene expression across 15 and 9 common cancer types, respectively. 70 significantly differentially expressed genes common to all 15 cancer types were enlisted. Correlating with protein expression levels from Human Protein Atlas, we observed 34 positively correlated gene sets which are enriched in gene expression, transcription from RNA Pol-II, regulation of transcription and mitotic cell cycle biological processes. Further, 24 lncRNAs were among common significantly differentially expressed non-coding genes. Using guilt-by-association method, we predicted lncRNAs to be involved in same biological processes. Combining RNA-RNA interaction prediction and transcription regulatory networks, we identified E2F1, FOXM1 and PVT1 regulatory path as recurring pan-cancer regulatory entity. PVT1 is predicted to interact with SYNE1 at 3′-UTR; DNAJC9, RNPS1 at 5′-UTR and ATXN2L, ALAD, FOXM1 and IRAK1 at CDS sites. The key findings are that through E2F1, FOXM1 and PVT1 regulatory axis and possible interactions with different coding genes, PVT1 may be playing a prominent role in pan-cancer development and progression.

An Induction System for Clustered Stomata by Sugar Solution Immersion Treatment in Arabidopsis thaliana Seedlings.

Stomatal movement mediates plant gas exchange, which is essential for photosynthesis and transpiration. Stomatal opening and closing are accomplished by a significant increase and decrease in guard cell volume, respectively. Because shuttle transport of ions and water occurs between guard cells and larger neighboring epidermal cells during stomatal movement, the spaced distribution of plant stomata is considered an optimal distribution for stomatal movement. Experimental systems for perturbing the spaced pattern of stomata are useful to examine the spacing pattern's significance. Several key genes associated with the spaced stomatal distribution have been identified, and clustered stomata can be experimentally induced by altering these genes. Alternatively, clustered stomata can be also induced by exogenous treatments without genetic modification. In this article, we describe a simple induction system for clustered stomata in Arabidopsis thaliana seedlings by immersion treatment with a sucrose-containing medium solution. Our method is easy and directly applicable to transgenic or mutant lines. Larger chloroplasts are presented as a cell biological hallmark of sucrose-induced clustered guard cells. In addition, a representative confocal microscopic image of cortical microtubules is shown as an example of intracellular observation of clustered guard cells. The radial orientation of cortical microtubules is maintained in clustered guard cells as in spaced guard cells in control conditions.

A Vault-Encapsulated Enzyme Approach for Efficient Degradation and Detoxification of Bisphenol A and Its Analogues.

We report an effective and environmentally sustainable water treatment approach using enzymes encapsulated in biogenic vault nanoparticles. Manganese peroxidase (MnP), whose stability was remarkably extended by encapsulating into vaults, rapidly catalyzed the biotransformation of endocrine-disrupting compounds, including bisphenol A (BPA), bisphenol F (BPF), and bisphenol AP (BPAP). The vault-encapsulated MnP (vMnP) treatment removed 80-95% of each of the tested bisphenols (BPs) at lower enzyme dosage, while free native MnP (nMnP) only resulted in a 19-36% removal, over a 24-h period. Treatment by vMnP and nMnP resulted in considerable disparities in product species and abundance, which were consistent with the observed changes in the estrogenic activities of BPs. To test if vMnP-catalyzed transformations generated toxic intermediates, we assessed biological hallmarks of BP toxicity, namely, the ability to disrupt reproductive processes. The toxicity of vMnP-treated samples, as measured in the model organism, Caenorhabditis elegans, was dramatically reduced for all three BPs, including the reproductive indicators of BPA exposure such as reduced fertility and increased germ cell death. Collectively, our results indicate that the vMnP system represents an efficient and safe approach for the removal of BPs and promise the development of vault-encapsulated customized enzymes for treating other targeted organic compounds in contaminated waters.

Single-cell sequencing in ovarian cancer: a new frontier in precision medicine.

This article discusses the advances, applications and challenges of using single-cell RNA sequencing data in guiding treatment decisions for ovarian cancer. Genetic heterogeneity is a hallmark of ovarian cancer biology and underlies treatment resistance. Defining the different cell types present within a single ovarian cancer is difficult, but could ultimately lead to improvements in diagnosis and treatment. Next-generation sequencing technologies have rapidly increased our understanding of the molecular landscape of epithelial ovarian cancers, but the majority of these studies are conducted on bulk samples, resulting in data that represents an 'average' of all cells present. Single-cell sequencing provides a means to characterize heterogeneity with a tumor tissue in ovarian cancer patients and opens up opportunity to determine key molecular properties that influence clinical outcomes, including prognosis and treatment response. Single-cell sequencing provides a powerful tool in improving our understanding of tumor cell heterogeneity for the purpose of informing personalized cancer treatment.

Blocking OLFM4/HIF-1α axis alleviates hypoxia-induced invasion, epithelial-mesenchymal transition, and chemotherapy resistance in non-small-cell lung cancer.

Hypoxia is a common biological hallmark of solid cancers, which has been proposed to be associated with oncogenesis and chemotherapy resistance. The purpose of the present study was to investigate the role and underlying mechanisms of olfactomedin 4 (OLFM4) in the hypoxia-induced invasion, epithelial-mesenchymal transition (EMT), and chemotherapy resistance of non-small-cell lung cancer (NSCLC). We observed dramatically upregulated expression of OLFM4 in several NSCLC cell lines, and this effect was more pronounced in A549 and H1299 cells. In addition, our data revealed that OLFM4 expression was remarkably increased in both A549 and H1299 cells under hypoxic microenvironment, accompanied by enhanced levels of hypoxia-inducible factor (HIF)-1α protein. The HIF-1α level was elevated in response to hypoxia, resulting in the regulation of OLFM4. Interestingly, OLFM4 was a positive regulator of hypoxia-driven HIF-1α production. Moreover, depletion of OLFM4 modulated multiple EMT-associated proteins, as evidenced by the enhanced E-cadherin levels along with the diminished expression of N-cadherin and vimentin in response to hypoxia, and thus blocked invasion ability of A549 and H1299 cells following exposure to hypoxia. Furthermore, ablation of OLFM4 accelerated the sensitivity of A549 cells to cisplatin under hypoxic conditions, implying that OLFM4 serves as a key regulator in chemotherapeutic resistance under hypoxia. In conclusion, OLFM4/HIF-1α axis might be a potential therapeutic strategy for NSCLC.

Is blood cortisol or vateritic otolith composition associated with natal dispersal or reproductive performance on the spawning grounds of straying and homing hatchery-produced chum salmon (Oncorhynchus keta) in Southeast Alaska?

ABSTRACTHoming with high fidelity to natal spawning grounds for reproduction is a hallmark of anadromous Pacific salmon biology, although low rates of dispersal (‘straying’) also occurs. Currently little is known about the proximate factors influencing straying, which limits our understanding of this fundamental biological phenomenon and impedes options for reducing straying-mediated interactions between wild and hatchery-produced individuals. We explored the potential role of stress experienced in captivity prior to intentional release to manifest in developmental irregularities and potentially influence rates of straying by adults. We compared two proxies for stress between groups of hatchery-produced individuals that had homed back to the hatchery or strayed to non-natal streams compared to wild individuals that were presumed to have homed to a wild spawning stream. Blood plasma cortisol was used to assess stress at the terminus of their migration, and percent frequency of vateritic otolith development within groups as a measure of stresses incurred during development. We found no evidence that either proxy for stress was associated with straying. No differences in cortisol concentrations were found between wild and hatchery-produced chum salmon that had homed or strayed, either in males (wild=95.9±175.7 ng/ml; stray=113.4±99.7 ng/ml; home=124.7±113.8 ng/ml) or females (wild=307.6±83.4 ng/ml; stray= 329.0±208.9 ng/ml; home=294.1±134.8 ng/ml); however, significant differences between males and females occurred in each group. The percent frequency of vaterite occurrence in otoliths of hatchery-produced chum salmon that either strayed (40% vaterite) or homed (45% vaterite) did not differ significantly, though rates of vaterite occurred less frequently in wild chum salmon (24%), which is consistent with other studies. Mass thermal marking of juvenile fish in hatcheries is unlikely to increase vateritic development as neither intensity (number of temperature changes) or complexity (number of temperature change sequences) of the mark was associated with frequency of vaterite occurrence. Though not associated with straying, cortisol concentrations were associated with shorter instream lifespan of both hatchery and wild individuals but did not appear to influence rates of egg retention in spawning females, suggesting an equivocal role in reproductive ecology. Our results are suggestive that stress induced during the early stages of rearing in a hatchery environment from marking or other causes may not increase straying later in life, though the higher rates of vaterite observed in hatchery-produced fish may come at a cost of increased marine mortality, due to the otoliths' role in navigation and hearing.

Down Syndrome, Ageing and Epigenetics.

During the past decades, life expectancy of subjects with Down syndrome (DS) has greatly improved, but age-specific mortality rates are still important and DS subjects are characterized by an acceleration of the ageing process, which affects particularly the immune and central nervous systems. In this chapter, we will first review the characteristics of the ageing phenomenon in brain and in immune system in DS and we will then discuss the biological hallmarks of ageing in this specific population. Finally, we will also consider in detail the knowledge on epigenetics in DS, particularly DNA methylation.

Chapter 23 - Clinical neurophysiology of CNS hypersomnias

Central nervous system hypersomnias (narcolepsy type 1 and type 2, idiopathic hypersomnia, and Kleine-Levin syndrome) are orphan sleep disorders in which the predominant symptom is excessive daytime sleepiness. The evaluation of sleepiness requires rigorous clinical and neurophysiologic approaches that may include the Epworth Sleepiness Scale, multiple sleep latency tests, and the maintenance of wakefulness test. However, to date, no gold standard measurement of excessive sleepiness exists, and there are no quantifiable biologic markers. The main pathophysiologic feature of central hypersomnias is thought to reflect a deficiency of arousal systems, rather than an overactivity of sleep systems or an imbalance between those systems. Impaired neurotransmission of hypocretin/orexin (neuropeptides of the lateral hypothalamus) is involved in the neurobiology of narcolepsy with cataplexy (NT1). NT1 is a well-characterized disorder, due to the destruction of hypocretin/orexin neurons by a probable autoimmune process. The biologic hallmarks of the other central hypersomnias remain unknown, and neurophysiologic biomarkers are still of major importance for the diagnosis and characterization of those disorders.

Therapeutic Manipulation of Cancer Cell Metabolism with the Mitochondrial Metabolism Inhibitor Cpi-613 in Addition to Chemotherapy Abrogates the Adverse Prognostic Effect of Age in Relapsed and Refractory AML

Background: Acute myeloid leukemia is an aggressive malignancy with poor outcomes especially in patients 60 years of age or older. This thought to be in part from increased resistance to chemotherapy in AML cells arising in an older host. One of the nine recognized biological hallmarks of aging is a decline in mitochondrial quality. The effect of exploiting this age-related metabolic vulnerability in relapsed AML has not been previously established. CPI-613 is a first-in-class agent that inhibits pyruvate dehydrogenase and α-ketoglutarate dehydrogenase, effectively impairing the TCA cycle. We have combined CPI-613 with high dose cytarabine and mitoxantrone in phase I and II clinical trials in over 100 patients with relapsed or refractory AML.Methods: To determine the effect of the addition of CPI-613 in older patients, the phase I and II datasets were combined and outcomes by age were analyzed and compared to age related outcomes in a historical dataset of patients treated with a high dose cytarabine, mitoxantrone and L-asparaginase but without CPI-613. In both trials, patients were given CPI-613 as a 2-hour infusion on days 1 through 5. Cytarabine was dosed at 3,000 mg/m² (if younger than 60) or at 1,500 mg/m² (if 60 years of age or older), given every 12 hours for 5 doses, starting on day 3 following the CPI-613 infusion. The mitoxantrone was dosed at 6 mg/m² and is given once daily following the first, third and fifth cytarabine doses. At day 14, nadir marrow was evaluated, and patients with residual disease could be re-treated as above or with an abbreviated 3-day cycle. Responding patients were eligible to receive up to 2 abbreviated 3-day consolidation cycles. The historical cohort was treated identically without CPI-613, except L-Asparaginase was given at a dose of 6,000 units/m2 following the last dose of cytarabine.Results: Patient characteristics are summarized in Table 1. In the historical dataset younger patients had a highly significant increase in median overall survival when compared to patients ≥60 years of age (figure 1A). In contrast, older and younger patients treated with CPI-613 in addition to the chemotherapy had no significant difference in median survival (figure 1B). Additionally, when outcome by dose of CPI-613 was analyzed, older but not younger patients had a significant improvement in survival when given a dose of 2,000 mg/m2 compared to those given 1,500 mg/m2 (figure 1C+D). Patients 60 years of age or older had a response rate of 55% and a median overall survival of 12.4 months when treated with a dose of CPI-613 of 2,000 mg/m2.Conclusions: Targeting mitochondrial metabolism exploits an age-related metabolic vulnerability in AML arising in older patients. These results have led to a randomized phase III trial of CPI-613 at 2,000 mg/m2 in combination with high dose cytarabine and mitoxantrone compared to high dose cytarabine and mitoxantrone alone in relapsed or refractory AML patients 60 years of age or older. [Display omitted] DisclosuresPardee:Amgen: Speakers Bureau; Karyopharm: Research Funding; Celgene: Speakers Bureau; Rafael Pharmaceuticals: Employment; Novartis: Speakers Bureau. Ellis:Alexion: Speakers Bureau. Powell:Rafael Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees.

Turning foes to friends: targeting cancer-associated fibroblasts.

Current paradigms of cancer-centric therapeutics are usually not sufficient to eradicate the malignancy, as the cancer stroma may prompt tumour relapse and therapeutic resistance. Among all the stromal cells that populate the tumour microenvironment, cancer-associated fibroblasts (CAFs) are the most abundant and are critically involved in cancer progression. CAFs regulate the biology of tumour cells and other stromal cells via cell-cell contact, releasing numerous regulatory factors and synthesizing and remodelling the extracellular matrix, and thus these cells affect cancer initiation and development. The recent characterization of CAFs based on specific cell surface markers not only deepens our insight into their phenotypic heterogeneity and functional diversity but also brings CAF-targeting therapies for cancer treatment onto the agenda. In this Review, we discuss the current knowledge of biological hallmarks, cellular origins, phenotypical plasticity and functional heterogeneity of CAFs and underscore their contribution to cancer progression. Moreover, we highlight relevant translational advances and potential therapeutic strategies that target CAFs for cancer treatment.