bims-pideca 2019-12-15 papers

bims-pideca

Biomed News

on Class IA PI3K signalling in development and cancer

Issue of 2019–12–15
25 papers selected by
Ralitsa Radostinova Madsen, University College London

Targeting mTOR and Metabolism in Cancer: Lessons and Innovations.
Phosphatidylinositol 3,4-bisphosphate synthesis and turnover are spatially segregated in the endocytic pathway.
Torin2 Exploits Replication and Checkpoint Vulnerabilities to Cause Death of PI3K-Activated Triple-Negative Breast Cancer Cells.
PTEN: Bridging Endocytosis and Signaling.
Defining How Oncogenic and Developmental Mutations of PIK3R1 Alter the Regulation of Class IA Phosphoinositide 3-Kinases.
PI3K Inhibitors in Breast Cancer Therapy.
Disruption of Acetyl-Lysine Turnover in Muscle Mitochondria Promotes Insulin Resistance and Redox Stress without Overt Respiratory Dysfunction.
Insight into the mechanism of allosteric activation of PI3Kα by oncoprotein K-Ras4B.
KRAS4A directly regulates hexokinase 1.
Phase 1 Dose Escalation Study of the Allosteric AKT Inhibitor BAY 1125976 in Advanced Solid Cancer-Lack of Association between Activating AKT Mutation and AKT Inhibition-Derived Efficacy.
Greasing the Wheels of the Cancer Machine: The Role of Lipid Metabolism in Cancer.
Pan-cancer molecular subtypes revealed by mass-spectrometry-based proteomic characterization of more than 500 human cancers.
Reprogramming of fatty acid metabolism in cancer.
Genetic determinants of the molecular portraits of epithelial cancers.
Mathematical Models of Cancer Cell Plasticity.
p27 allosterically activates cyclin-dependent kinase 4 and antagonizes palbociclib inhibition.
Endogenous IGF Signaling Directs Heterogeneous Mesoderm Differentiation in Human Embryonic Stem Cells.
Rapamycin persistently improves cardiac function in aged, male and female mice, even following cessation of treatment.
The functional landscape of the human phosphoproteome.
Transcriptomic analysis of human primary breast cancer identifies fatty acid oxidation as a target for metformin.
ATP citrate lyase: a central metabolic enzyme in cancer.
Histone proteomics reveals novel post-translational modifications in breast cancer.
Utility of Reverse-Phase Protein Array for Refining Precision Oncology.
The Metabolic Interplay between Cancer and Other Diseases.
Autophagy in cancer: moving from understanding mechanism to improving therapy responses in patients.

Cells. 2019 Dec 06. pii: E1584. [Epub ahead of print]8(12):

Targeting mTOR and Metabolism in Cancer: Lessons and Innovations.

Cedric Magaway, Eugene Kim, Estela Jacinto.

  Cancer cells support their growth and proliferation by reprogramming their metabolism in order to gain access to nutrients. Despite the heterogeneity in genetic mutations that lead to tumorigenesis, a common alteration in tumors occurs in pathways that upregulate nutrient acquisition. A central signaling pathway that controls metabolic processes is the mTOR pathway. The elucidation of the regulation and functions of mTOR can be traced to the discovery of the natural compound, rapamycin. Studies using rapamycin have unraveled the role of mTOR in the control of cell growth and metabolism. By sensing the intracellular nutrient status, mTOR orchestrates metabolic reprogramming by controlling nutrient uptake and flux through various metabolic pathways. The central role of mTOR in metabolic rewiring makes it a promising target for cancer therapy. Numerous clinical trials are ongoing to evaluate the efficacy of mTOR inhibition for cancer treatment. Rapamycin analogs have been approved to treat specific types of cancer. Since rapamycin does not fully inhibit mTOR activity, new compounds have been engineered to inhibit the catalytic activity of mTOR to more potently block its functions. Despite highly promising pre-clinical studies, early clinical trial results of these second generation mTOR inhibitors revealed increased toxicity and modest antitumor activity. The plasticity of metabolic processes and seemingly enormous capacity of malignant cells to salvage nutrients through various mechanisms make cancer therapy extremely challenging. Therefore, identifying metabolic vulnerabilities in different types of tumors would present opportunities for rational therapeutic strategies. Understanding how the different sources of nutrients are metabolized not just by the growing tumor but also by other cells from the microenvironment, in particular, immune cells, will also facilitate the design of more sophisticated and effective therapeutic regimen. In this review, we discuss the functions of mTOR in cancer metabolism that have been illuminated from pre-clinical studies. We then review key findings from clinical trials that target mTOR and the lessons we have learned from both pre-clinical and clinical studies that could provide insights on innovative therapeutic strategies, including immunotherapy to target mTOR signaling and the metabolic network in cancer.

Keywords:  cancer metabolism; kinase inhibitors; mTOR in immunotherapy; mTOR inhibitors; mTOR signaling; mTORC1; mTORC2; metabolism; nutrient metabolism; rapalogs

DOI:  https://doi.org/10.3390/cells8121584
J Biol Chem. 2019 Dec 12. pii: jbc.RA119.011774. [Epub ahead of print]

Phosphatidylinositol 3,4-bisphosphate synthesis and turnover are spatially segregated in the endocytic pathway.

Haibin Wang, Dinah Loerke, Caroline Bruns, Rainer Müller, Philipp Alexander Koch, Dmytro Puchkov, Carsten Schultz, Volker Haucke.

  Phosphoinositides play crucial roles in intracellular membrane dynamics and cell signaling, with phosphatidylinositol (PI) 3-phosphates being the predominant phosphoinositide lipids at endosomes and lysosomes, whereas PI 4-phosphates such as phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] are enriched at the cell surface including sites of endocytosis. How PI 4-phosphates and PI 3-phosphates are dynamically interconverted within the endocytic pathway and how this is controlled in space and time remains poorly understood. Here, combining live imaging, genome engineering, and acute chemical and genetic manipulations, we found that local synthesis of PI(3,4)P2 by phosphatidylinositol 3-kinase C2α (PI3KC2α) at plasma membrane clathrin-coated pits (CCPs) is spatially segregated from its hydrolysis by the PI(3,4)P2-specific inositol polyphosphate 4-phosphatase 4A (INPP4A). We observed that INPP4A is dispensable for clathrin-mediated endocytosis (CME) and is undetectable in endocytic CCPs. Instead, we find that INPP4A partially localizes to endosomes and that loss of INPP4A in HAP1 cancer cells perturbs signaling via AKT kinase (AKT) and mTOR complex 1 (mTORC1). These results reveal a function for INPP4-mediated PI(3,4)P2 hydrolysis in the local regulation of growth factor and nutrient signals at endosomes in cancer cells. They further suggest a model according to which the synthesis and turnover of PI(3,4)P2 are spatially segregated within the endocytic pathway to couple endocytic membrane traffic to growth factor and nutrient signaling.

Keywords:  clathrin; endocytosis; endosome; imaging; lysosome; mTOR complex (mTORC); nutrient signaling; phosphatidylinositol kinase (PI Kinase); phosphatidylinositol phosphatase; phosphoinositide

DOI:  https://doi.org/10.1074/jbc.RA119.011774
Cell Syst. 2019 Dec 02. pii: S2405-4712(19)30387-4. [Epub ahead of print]

Torin2 Exploits Replication and Checkpoint Vulnerabilities to Cause Death of PI3K-Activated Triple-Negative Breast Cancer Cells.

Sameer S Chopra, Anne Jenney, Adam Palmer, Mario Niepel, Mirra Chung, Caitlin Mills, Sindhu Carmen Sivakumaren, Qingsong Liu, Jia-Yun Chen, Clarence Yapp, John M Asara, Nathanael S Gray, Peter K Sorger.

  Frequent mutation of PI3K/AKT/mTOR signaling pathway genes in human cancers has stimulated large investments in targeted drugs but clinical successes are rare. As a result, many cancers with high PI3K pathway activity, such as triple-negative breast cancer (TNBC), are treated primarily with chemotherapy. By systematically analyzing responses of TNBC cells to a diverse collection of PI3K pathway inhibitors, we find that one drug, Torin2, is unusually effective because it inhibits both mTOR and other PI3K-like kinases (PIKKs). In contrast to mTOR-selective inhibitors, Torin2 exploits dependencies on several kinases for S-phase progression and cell-cycle checkpoints, thereby causing accumulation of single-stranded DNA and death by replication catastrophe or mitotic failure. Thus, Torin2 and its chemical analogs represent a mechanistically distinct class of PI3K pathway inhibitors that are uniquely cytotoxic to TNBC cells. This insight could be translated therapeutically by further developing Torin2 analogs or combinations of existing mTOR and PIKK inhibitors.

Keywords:  ATR/Chk1; PI3K/AKT/mTOR; Torin2; cell cycle; mitotic catastrophe; polypharmacology; replication catastrophe; replication stress; small molecule drugs; triple-negative breast cancer

DOI:  https://doi.org/10.1016/j.cels.2019.11.001
Cold Spring Harb Perspect Med. 2019 Dec 09. pii: a036103. [Epub ahead of print]

PTEN: Bridging Endocytosis and Signaling.

Matthew F Lee, Lloyd C Trotman.

The transduction of signals in the PTEN/PI3-kinase (PI3K) pathway is built around a phosphoinositide (PIP) lipid messenger, phosphatidylinositol trisphosphate, PI(3,4,5)P3 or PIP3 Another, more ancient role of this family of messengers is the control of endocytosis, where a handful of separate PIPs act like postal codes. Prominent among them is PI(3)P, which helps to ensure that endocytic vesicles, their cargo, and membranes themselves reach their correct destinations. Traditionally, the cancer and the endocytic functions of the PI3K signaling pathway have been studied by cancer and membrane biologists, respectively, with some notable but overall minimal overlap. Modern microscopy has enabled monitoring of the PTEN/PI3K pathway in action. Here, we explore the flurry of groundbreaking concepts emerging from those efforts. The discovery that PTEN contains an autonomous PI(3)P reader domain, fused to the catalytic PIP3 eraser domain has prompted us to explore the relationship between PI3K signaling and endocytosis. This revealed how PTEN can achieve signal termination in a precisely controlled fashion, because endocytosis can package the PIP3 signal into discrete units that PTEN will erase. We explore how PTEN can bridge the worlds of endocytosis and PI3K signaling and discuss progress on how PI3K/AKT signaling can be acting from internal membranes. We discuss how the PTEN/PI3K system for growth control may have emerged from principles of endocytosis, and how this development could have affected the evolution of multicellular organisms.

DOI: https://doi.org/10.1101/cshperspect.a036103
Structure. 2019 Nov 29. pii: S0969-2126(19)30397-1. [Epub ahead of print]

Defining How Oncogenic and Developmental Mutations of PIK3R1 Alter the Regulation of Class IA Phosphoinositide 3-Kinases.

Gillian L Dornan, Jordan T B Stariha, Manoj K Rathinaswamy, Cameron J Powell, Martin J Boulanger, John E Burke.

  The class I phosphoinositide 3-kinases (PI3Ks) are key signaling enzymes composed of a heterodimer of a p110 catalytic subunit and a p85 regulatory subunit, with PI3K mutations being causative of multiple human diseases including cancer, primary immunodeficiencies, and developmental disorders. Mutations in the p85α regulatory subunit encoded by PIK3R1 can both activate PI3K through oncogenic truncations in the iSH2 domain, or inhibit PI3K through developmental disorder mutations in the cSH2 domain. Using a combined biochemical and hydrogen deuterium exchange mass spectrometry approach we have defined the molecular basis for how these mutations alter the activity of p110α/p110δ catalytic subunits. We find that the oncogenic Q572∗ truncation of PIK3R1 disrupts all p85-inhibitory inputs, with p110α being hyper-activated compared with p110δ. In addition, we find that the R649W mutation in the cSH2 of PIK3R1 decreases sensitivity to activation by receptor tyrosine kinases. This work reveals unique insight into isoform-specific regulation of p110s by p85α.

Keywords:  HDX-MS; PI3K; PI3K-Akt; PIK3CA; PIK3R1; hydrogen exchange; p110; p85; phosphoinositide 3-kinase; phosphoinositides

DOI:  https://doi.org/10.1016/j.str.2019.11.013
Curr Oncol Rep. 2019 Dec 11. 21(12): 110

PI3K Inhibitors in Breast Cancer Therapy.

Haley Ellis, Cynthia X Ma.

   PURPOSE OF REVIEW: The phosphatidylinositol 3-kinase (PI3K) pathway is the most common aberrantly activated pathway in breast cancer, making it an attractive therapeutic target. In this review, we will discuss the rationale for targeting PI3K/AKT signaling and the development of PI3K/AKT inhibitors in breast cancer.
RECENT FINDINGS: Although the initial clinical trials with pan-PI3K inhibitors were challenged by high toxicities and modest antitumor effect, there has been continued effort to develop agents more precisely targeting PI3K isoforms to improve therapeutic index. Alpelisib in combination with fulvestrant is now available in the clinic for postmenopausal women with advanced or metastatic hormone receptor (HR)-positive, HER2-negative, PIK3CA-mutated breast cancer. In addition, promising data has been observed in randomized phase II trials of AKT inhibitors in combination with fulvestrant or paclitaxel in metastatic HR-positive, HER2-negative disease and triple negative breast cancer (TNBC), respectively. The high frequency of genetic alterations in the PI3K pathway has provided the rationale for development of inhibitors targeting PI3K/AKT. Despite initial disappointment with several randomized trials of pan-PI3K inhibitors in HR-positive breast cancer, there has been continued effort to more precisely target PI3K isoforms, which has led to clinical benefit for patients with advanced breast cancer.

Keywords:  AKT inhibitor; Breast cancer; PI3K inhibitor; PI3K pathway; Targeted therapy

DOI:  https://doi.org/10.1007/s11912-019-0846-7
Cell Metab. 2019 Nov 28. pii: S1550-4131(19)30610-2. [Epub ahead of print]

Disruption of Acetyl-Lysine Turnover in Muscle Mitochondria Promotes Insulin Resistance and Redox Stress without Overt Respiratory Dysfunction.

Ashley S Williams, Timothy R Koves, Michael T Davidson, Scott B Crown, Kelsey H Fisher-Wellman, Maria J Torres, James A Draper, Tara M Narowski, Dorothy H Slentz, Louise Lantier, David H Wasserman, Paul A Grimsrud, Deborah M Muoio.

  This study sought to examine the functional significance of mitochondrial protein acetylation using a double knockout (DKO) mouse model harboring muscle-specific deficits in acetyl-CoA buffering and lysine deacetylation, due to genetic ablation of carnitine acetyltransferase and Sirtuin 3, respectively. DKO mice are highly susceptible to extreme hyperacetylation of the mitochondrial proteome and develop a more severe form of diet-induced insulin resistance than either single KO mouse line. However, the functional phenotype of hyperacetylated DKO mitochondria is largely normal. Of the >120 measures of respiratory function assayed, the most consistently observed traits of a markedly heightened acetyl-lysine landscape are enhanced oxygen flux in the context of fatty acid fuel and elevated rates of electron leak. In sum, the findings challenge the notion that lysine acetylation causes broad-ranging damage to mitochondrial quality and performance and raise the possibility that acetyl-lysine turnover, rather than acetyl-lysine stoichiometry, modulates redox balance and carbon flux.

Keywords:  NAD biology; bioenergetics; diabetes; fat oxidation; fatty acid oxidation; insulin action; lysine acetylation; mitochondria; muscle; nutrition; obesity; proteomics; reactive oxygen species; redox; respiration; sirtuins

DOI:  https://doi.org/10.1016/j.cmet.2019.11.003
Int J Biol Macromol. 2019 Dec 06. pii: S0141-8130(19)37606-8. [Epub ahead of print]

Insight into the mechanism of allosteric activation of PI3Kα by oncoprotein K-Ras4B.

Xinyi Li, Jinyuan Dai, Duan Ni, Xinheng He, Hao Zhang, Jian Zhang, Qiang Fu, Yaqin Liu, Shaoyong Lu.

  Ras is a key member in the superfamily of small GTPase. Transforming between GTP-bound active state and GDP-bound inactive state in response to exogenous signals, Ras serves as a binary switch in various signaling pathways. One of its downstream effectors is phosphatidylinositol-4,5-bisphosphate 3-kinase α (PI3Kα), which phosphorylates phosphatidylinositol 4,5-bisphosphate into phosphatidylinositol 3,4,5-trisphosphate in the PI3K/Akt/mTOR pathway and mediates an array of important cellular activities including cell growth, migration and survival. Hyperactivation of PI3Kα induced by the Ras isoform K-Ras4B has been unveiled as a key event during the oncogenesis of pancreatic ductal adenocarcinoma, but the underlying mechanism of how K-Ras4B allosterically activates PI3Kα still remains largely unsolved. Here, we employed accelerated molecular dynamic simulations and allosteric pathway analysis to explore into the activation process of PI3Kα by K-Ras4B and unraveled the underlying structural mechanisms. We found that K-Ras4B binding induced more conformational dynamics within PI3Kα and triggered its step-wise transition from a self-inhibited state towards an activated state. Moreover, K-Ras4B binding markedly disrupted the interactions along the p110/p85 interface, especially the ones between nSH2 in p85 and its nearby functional domains in p110 like C2, helical, and kinase domains. The altered inter-domain interactions exposed the kinase domain, which promoted the membrane association and substrate phosphorylation of PI3Kα, thereby facilitating its activation. In particular, the community networks and allosteric pathways analysis further revealed that in PI3Kα/K-Ras4B system, allosteric signaling regulating p110/p85 interaction was rewired from the helical domain to the kinase domain and several important residues and their related allosteric pathways mediating PI3Kα autoinhibition were bypassed. The obtained structural mechanisms provide an in-depth mechanistic insight into the allosteric activation of PI3Kα by K-Ras4B as well as shed light on its drug discovery.

Keywords:  Allosteric communication; Allosteric effects; Allosteric regulation; Molecular dynamics simulations; PI3K; Protein-protein interactions; Ras

DOI:  https://doi.org/10.1016/j.ijbiomac.2019.12.020
Nature. 2019 Dec 11.

KRAS4A directly regulates hexokinase 1.

Caroline R Amendola, James P Mahaffey, Seth J Parker, Ian M Ahearn, Wei-Ching Chen, Mo Zhou, Helen Court, Jie Shi, Sebastian L Mendoza, Michael J Morten, Eli Rothenberg, Eyal Gottlieb, Youssef Z Wadghiri, Richard Possemato, Stevan R Hubbard, Allan Balmain, Alec C Kimmelman, Mark R Philips.

The most frequently mutated oncogene in cancer is KRAS, which uses alternative fourth exons to generate two gene products (KRAS4A and KRAS4B) that differ only in their C-terminal membrane-targeting region1. Because oncogenic mutations occur in exons 2 or 3, two constitutively active KRAS proteins-each capable of transforming cells-are encoded when KRAS is activated by mutation2. No functional distinctions among the splice variants have so far been established. Oncogenic KRAS alters the metabolism of tumour cells3 in several ways, including increased glucose uptake and glycolysis even in the presence of abundant oxygen4 (the Warburg effect). Whereas these metabolic effects of oncogenic KRAS have been explained by transcriptional upregulation of glucose transporters and glycolytic enzymes3-5, it is not known whether there is direct regulation of metabolic enzymes. Here we report a direct, GTP-dependent interaction between KRAS4A and hexokinase 1 (HK1) that alters the activity of the kinase, and thereby establish that HK1 is an effector of KRAS4A. This interaction is unique to KRAS4A because the palmitoylation-depalmitoylation cycle of this RAS isoform enables colocalization with HK1 on the outer mitochondrial membrane. The expression of KRAS4A in cancer may drive unique metabolic vulnerabilities that can be exploited therapeutically.

DOI: https://doi.org/10.1038/s41586-019-1832-9
Cancers (Basel). 2019 Dec 10. pii: E1987. [Epub ahead of print]11(12):

Phase 1 Dose Escalation Study of the Allosteric AKT Inhibitor BAY 1125976 in Advanced Solid Cancer-Lack of Association between Activating AKT Mutation and AKT Inhibition-Derived Efficacy.

Andreas Schneeweiss, Dagmar Hess, Markus Joerger, Andrea Varga, Stacy Moulder, Apostolia M Tsimberidou, Cynthia Ma, Sara A Hurvitz, Christine Rentzsch, Marion Rudolph, Silke Thiele, Oliver Boix, Gary Wilkinson, Eleni Lagkadinou, Matthias Ocker.

  This open-label, phase I first-in-human study (NCT01915576) of BAY 1125976, a highly specific and potent allosteric inhibitor of AKT1/2, aimed to evaluate the safety, pharmacokinetics, and maximum tolerated dose of BAY 1125976 in patients with advanced solid tumors. Oral dose escalation was investigated with a continuous once daily (QD) treatment (21 days/cycle) and a twice daily (BID) schedule. A dose expansion in 28 patients with hormone receptor-positive metastatic breast cancer, including nine patients harboring the AKT1E17K mutation, was performed at the recommended phase 2 dose (R2D) of 60 mg BID. Dose-limiting toxicities (Grades 3-4) were increased in transaminases, γ-glutamyltransferase (γ-GT), and alkaline phosphatase in four patients in both schedules and stomach pain in one patient. Of the 78 patients enrolled, one patient had a partial response, 30 had stable disease, and 38 had progressive disease. The clinical benefit rate was 27.9% among 43 patients treated at the R2D. AKT1E17K mutation status was not associated with tumor response. Genetic analyses revealed additional mutations that could promote tumor cell growth despite the inhibition of AKT1/2. BAY 1125976 was well tolerated and inhibited AKT1/2 signaling but did not lead to radiologic or clinical tumor responses. Thus, the refinement of a selection of biomarkers for AKT inhibitors is needed to improve their monotherapy activity.

Keywords:  AKT inhibitor; biomarker; breast cancer; pharmacokinetics; phase 1

DOI:  https://doi.org/10.3390/cancers11121987
Cell Metab. 2019 Dec 03. pii: S1550-4131(19)30617-5. [Epub ahead of print]

Greasing the Wheels of the Cancer Machine: The Role of Lipid Metabolism in Cancer.

Marteinn Thor Snaebjornsson, Sudha Janaki-Raman, Almut Schulze.

  Altered lipid metabolism is among the most prominent metabolic alterations in cancer. Enhanced synthesis or uptake of lipids contributes to rapid cancer cell growth and tumor formation. Lipids are a highly complex group of biomolecules that not only constitute the structural basis of biological membranes but also function as signaling molecules and an energy source. Here, we summarize recent evidence implicating altered lipid metabolism in different aspects of the cancer phenotype and discuss potential strategies by which targeting lipid metabolism could provide a therapeutic window for cancer treatment.

Keywords:  cancer; fatty acid desaturation; fatty acids; lipid metabolism; lipid remodeling; mevalonate

DOI:  https://doi.org/10.1016/j.cmet.2019.11.010
Nat Commun. 2019 Dec 12. 10(1): 5679

Pan-cancer molecular subtypes revealed by mass-spectrometry-based proteomic characterization of more than 500 human cancers.

Fengju Chen, Darshan S Chandrashekar, Sooryanarayana Varambally, Chad J Creighton.

Mass-spectrometry-based proteomic profiling of human cancers has the potential for pan-cancer analyses to identify molecular subtypes and associated pathway features that might be otherwise missed using transcriptomics. Here, we classify 532 cancers, representing six tissue-based types (breast, colon, ovarian, renal, uterine), into ten proteome-based, pan-cancer subtypes that cut across tumor lineages. The proteome-based subtypes are observable in external cancer proteomic datasets surveyed. Gene signatures of oncogenic or metabolic pathways can further distinguish between the subtypes. Two distinct subtypes both involve the immune system, one associated with the adaptive immune response and T-cell activation, and the other associated with the humoral immune response. Two additional subtypes each involve the tumor stroma, one of these including the collagen VI interacting network. Three additional proteome-based subtypes-respectively involving proteins related to Golgi apparatus, hemoglobin complex, and endoplasmic reticulum-were not reflected in previous transcriptomics analyses. A data portal is available at UALCAN website.

DOI: https://doi.org/10.1038/s41467-019-13528-0
Br J Cancer. 2019 Dec 10.

Reprogramming of fatty acid metabolism in cancer.

Nikos Koundouros, George Poulogiannis.

A common feature of cancer cells is their ability to rewire their metabolism to sustain the production of ATP and macromolecules needed for cell growth, division and survival. In particular, the importance of altered fatty acid metabolism in cancer has received renewed interest as, aside their principal role as structural components of the membrane matrix, they are important secondary messengers, and can also serve as fuel sources for energy production. In this review, we will examine the mechanisms through which cancer cells rewire their fatty acid metabolism with a focus on four main areas of research. (1) The role of de novo synthesis and exogenous uptake in the cellular pool of fatty acids. (2) The mechanisms through which molecular heterogeneity and oncogenic signal transduction pathways, such as PI3K-AKT-mTOR signalling, regulate fatty acid metabolism. (3) The role of fatty acids as essential mediators of cancer progression and metastasis, through remodelling of the tumour microenvironment. (4) Therapeutic strategies and considerations for successfully targeting fatty acid metabolism in cancer. Further research focusing on the complex interplay between oncogenic signalling and dysregulated fatty acid metabolism holds great promise to uncover novel metabolic vulnerabilities and improve the efficacy of targeted therapies.

DOI: https://doi.org/10.1038/s41416-019-0650-z
Nat Commun. 2019 Dec 11. 10(1): 5666

Genetic determinants of the molecular portraits of epithelial cancers.

Youli Xia, Cheng Fan, Katherine A Hoadley, Joel S Parker, Charles M Perou.

The ability to characterize and predict tumor phenotypes is crucial to precision medicine. In this study, we present an integrative computational approach using a genome-wide association analysis and an Elastic Net prediction method to analyze the relationship between DNA copy number alterations and an archive of gene expression signatures. Across breast cancers, we are able to quantitatively predict many gene signatures levels within individual tumors with high accuracy based upon DNA copy number features alone, including proliferation status and Estrogen-signaling pathway activity. We can also predict many other key phenotypes, including intrinsic molecular subtypes, estrogen receptor status, and TP53 mutation. This approach is also applied to TCGA Pan-Cancer, which identify repeatedly predictable signatures across tumor types including immune features in lung squamous and basal-like breast cancers. These Elastic Net DNA predictors could also be called from DNA-based gene panels, thus facilitating their use as biomarkers to guide therapeutic decision making.

DOI: https://doi.org/10.1038/s41467-019-13588-2
J Oncol. 2019 ;2019 2403483

Mathematical Models of Cancer Cell Plasticity.

Hasitha N Weerasinghe, Pamela M Burrage, Kevin Burrage, Dan V Nicolau.

Quantitative modelling is increasingly important in cancer research, helping to integrate myriad diverse experimental data into coherent pictures of the disease and able to discriminate between competing hypotheses or suggest specific experimental lines of enquiry and new approaches to therapy. Here, we review a diverse set of mathematical models of cancer cell plasticity (a process in which, through genetic and epigenetic changes, cancer cells survive in hostile environments and migrate to more favourable environments, respectively), tumour growth, and invasion. Quantitative models can help to elucidate the complex biological mechanisms of cancer cell plasticity. In this review, we discuss models of plasticity, tumour progression, and metastasis under three broadly conceived mathematical modelling techniques: discrete, continuum, and hybrid, each with advantages and disadvantages. An emerging theme from the predictions of many of these models is that cell escape from the tumour microenvironment (TME) is encouraged by a combination of physiological stress locally (e.g., hypoxia), external stresses (e.g., the presence of immune cells), and interactions with the extracellular matrix. We also discuss the value of mathematical modelling for understanding cancer more generally.

DOI: https://doi.org/10.1155/2019/2403483
Science. 2019 Dec 13. pii: eaaw2106. [Epub ahead of print]366(6471):

p27 allosterically activates cyclin-dependent kinase 4 and antagonizes palbociclib inhibition.

Keelan Z Guiley, Jack W Stevenson, Kevin Lou, Krister J Barkovich, Vishnu Kumarasamy, Tilini U Wijeratne, Katharine L Bunch, Sarvind Tripathi, Erik S Knudsen, Agnieszka K Witkiewicz, Kevan M Shokat, Seth M Rubin.

The p27 protein is a canonical negative regulator of cell proliferation and acts primarily by inhibiting cyclin-dependent kinases (CDKs). Under some circumstances, p27 is associated with active CDK4, but no mechanism for activation has been described. We found that p27, when phosphorylated by tyrosine kinases, allosterically activated CDK4 in complex with cyclin D1 (CDK4-CycD1). Structural and biochemical data revealed that binding of phosphorylated p27 (phosp27) to CDK4 altered the kinase adenosine triphosphate site to promote phosphorylation of the retinoblastoma tumor suppressor protein (Rb) and other substrates. Surprisingly, purified and endogenous phosp27-CDK4-CycD1 complexes were insensitive to the CDK4-targeting drug palbociclib. Palbociclib instead primarily targeted monomeric CDK4 and CDK6 (CDK4/6) in breast tumor cells. Our data characterize phosp27-CDK4-CycD1 as an active Rb kinase that is refractory to clinically relevant CDK4/6 inhibitors.

DOI: https://doi.org/10.1126/science.aaw2106
Cell Rep. 2019 Dec 10. pii: S2211-1247(19)31529-3. [Epub ahead of print]29(11): 3374-3384.e5

Endogenous IGF Signaling Directs Heterogeneous Mesoderm Differentiation in Human Embryonic Stem Cells.

Yang Yang, Zhili Ren, Faxiang Xu, Ya Meng, Yumeng Zhang, Nana Ai, Yan Long, Hio Ian Fok, Chunhao Deng, Xianyang Zhao, Liancheng Huang, Qi Zhao, Jiaxian Wang, Weiwei Liu, Wei Ge, Guokai Chen.

  During embryogenesis, various cell types emerge simultaneously from their common progenitors under the influence of intrinsic signals. Human embryonic stem cells can differentiate to diverse cell types of three embryonic lineages, making them an excellent system for understanding the regulatory mechanism that maintains the balance of different cell types in embryogenesis. In this report, we demonstrate that insulin-like growth factor (IGF) proteins are endogenously expressed during differentiation, and their temporal expression contributes to the cell fate diversity in mesoderm differentiation. Small molecule LY294002 inhibits the IGF pathway to promote cardiomyocyte differentiation while suppressing epicardial and noncardiac cell fates. LY294002-induced cardiomyocytes demonstrate characteristic cardiomyocyte features and provide insights into the molecular mechanisms underlying cardiac differentiation. We further show that LY294002 induces cardiomyocytes through CK2 pathway inhibition. This study elucidates the crucial roles of endogenous IGF in mesoderm differentiation and shows that the inhibition of the IGF pathway is an effective approach for generating cardiomyocytes.

Keywords:  IGF; PI3K; WNT; cardiomyocyte; casein kinase II; heterogeneity; human embryonic stem cells; insulin; mesoderm; temporal regulation

DOI:  https://doi.org/10.1016/j.celrep.2019.11.047
Aging Cell. 2019 Dec 10. e13086

Rapamycin persistently improves cardiac function in aged, male and female mice, even following cessation of treatment.

Ellen Quarles, Nathan Basisty, Ying Ann Chiao, Gennifer Merrihew, Haiwei Gu, Mariya T Sweetwyne, Jeanne Fredrickson, Ngoc-Han Nguyen, Maria Razumova, Kristina Kooiker, Farid Moussavi-Harami, Michael Regnier, Christopher Quarles, Michael MacCoss, Peter S Rabinovitch.

  Even in healthy aging, cardiac morbidity and mortality increase with age in both mice and humans. These effects include a decline in diastolic function, left ventricular hypertrophy, metabolic substrate shifts, and alterations in the cardiac proteome. Previous work from our laboratory indicated that short-term (10-week) treatment with rapamycin, an mTORC1 inhibitor, improved measures of these age-related changes. In this report, we demonstrate that the rapamycin-dependent improvement of diastolic function is highly persistent, while decreases in both cardiac hypertrophy and passive stiffness are substantially persistent 8 weeks after cessation of an 8-week treatment of rapamycin in both male and female 22- to 24-month-old C57BL/6NIA mice. The proteomic and metabolomic abundance changes that occur after 8 weeks of rapamycin treatment have varying persistence after 8 further weeks without the drug. However, rapamycin did lead to a persistent increase in abundance of electron transport chain (ETC) complex components, most of which belonged to Complex I. Although ETC protein abundance and Complex I activity were each differentially affected in males and females, the ratio of Complex I activity to Complex I protein abundance was equally and persistently reduced after rapamycin treatment in both sexes. Thus, rapamycin treatment in the aged mice persistently improved diastolic function and myocardial stiffness, persistently altered the cardiac proteome in the absence of persistent metabolic changes, and led to persistent alterations in mitochondrial respiratory chain activity. These observations suggest that an optimal translational regimen for rapamycin therapy that promotes enhancement of healthspan may involve intermittent short-term treatments.

Keywords:  aging; echocardiography; heart; persistence; proteomics; rapamycin

DOI:  https://doi.org/10.1111/acel.13086
Nat Biotechnol. 2019 Dec 09.

The functional landscape of the human phosphoproteome.

David Ochoa, Andrew F Jarnuczak, Cristina Viéitez, Maja Gehre, Margaret Soucheray, André Mateus, Askar A Kleefeldt, Anthony Hill, Luz Garcia-Alonso, Frank Stein, Nevan J Krogan, Mikhail M Savitski, Danielle L Swaney, Juan A Vizcaíno, Kyung-Min Noh, Pedro Beltrao.

Protein phosphorylation is a key post-translational modification regulating protein function in almost all cellular processes. Although tens of thousands of phosphorylation sites have been identified in human cells, approaches to determine the functional importance of each phosphosite are lacking. Here, we manually curated 112 datasets of phospho-enriched proteins, generated from 104 different human cell types or tissues. We re-analyzed the 6,801 proteomics experiments that passed our quality control criteria, creating a reference phosphoproteome containing 119,809 human phosphosites. To prioritize functional sites, we used machine learning to identify 59 features indicative of proteomic, structural, regulatory or evolutionary relevance and integrate them into a single functional score. Our approach identifies regulatory phosphosites across different molecular mechanisms, processes and diseases, and reveals genetic susceptibilities at a genomic scale. Several regulatory phosphosites were experimentally validated, including identifying a role in neuronal differentiation for phosphosites in SMARCC2, a member of the SWI/SNF chromatin-remodeling complex.

DOI: https://doi.org/10.1038/s41587-019-0344-3
Br J Cancer. 2019 Dec 10.

Transcriptomic analysis of human primary breast cancer identifies fatty acid oxidation as a target for metformin.

Simon R Lord, Jennifer M Collins, Wei-Chen Cheng, Syed Haider, Simon Wigfield, Edoardo Gaude, Barbara A Fielding, Katherine E Pinnick, Ulrike Harjes, Ashvina Segaran, Pooja Jha, Gerald Hoefler, Michael N Pollak, Alastair M Thompson, Pankaj G Roy, Ruth English, Rosie F Adams, Christian Frezza, Francesca M Buffa, Fredrik Karpe, Adrian L Harris.

BACKGROUND: Epidemiological studies suggest that metformin may reduce the incidence of cancer in patients with diabetes and multiple late phase clinical trials assessing the potential of repurposing this drug are underway. Transcriptomic profiling of tumour samples is an excellent tool to understand drug bioactivity, identify candidate biomarkers and assess for mechanisms of resistance to therapy.
METHODS: Thirty-six patients with untreated primary breast cancer were recruited to a window study and transcriptomic profiling of tumour samples carried out before and after metformin treatment.
RESULTS: Multiple genes that regulate fatty acid oxidation were upregulated at the transcriptomic level and there was a differential change in expression between two previously identified cohorts of patients with distinct metabolic responses. Increase in expression of a mitochondrial fatty oxidation gene composite signature correlated with change in a proliferation gene signature. In vitro assays showed that, in contrast to previous studies in models of normal cells, metformin reduces fatty acid oxidation with a subsequent accumulation of intracellular triglyceride, independent of AMPK activation.
CONCLUSIONS: We propose that metformin at clinical doses targets fatty acid oxidation in cancer cells with implications for patient selection and drug combinations.
CLINICAL TRIAL REGISTRATION: NCT01266486.

DOI: https://doi.org/10.1038/s41416-019-0665-5
Cancer Lett. 2019 Dec 09. pii: S0304-3835(19)30616-0. [Epub ahead of print]

ATP citrate lyase: a central metabolic enzyme in cancer.

Philippe Icard, Zherui Wu, Ludovic Fournel, Antoine Coquerel, Hubert Lincet, Marco Alifano.

  ACLY links energy metabolism provided by catabolic pathways to biosynthesis. ACLY, which has been found to be overexpressed in many cancers, converts citrate into acetyl-CoA and OAA. The first of these moleculessupportsprotein acetylation, in particular that of histone, and de novo lipid synthesis, and the last one sustains the production of aspartate (required for nucleotide and polyamine synthesis) and the regeneration of NADPH,H+(consumed in redox reaction and biosynthesis). ACLY transcription is promoted by SREBP1, its activity is stabilized by acetylation and promoted by AKT phosphorylation (stimulated by growth factors and glucose abundance). ACLY plays a pivotal role in cancer metabolism through the potential deprivation of cytosolic citrate, a process promoting glycolysis through the enhancement of the activities of PFK 1 and 2 with concomitant activation of oncogenic drivers such as PI3K/AKT which activate ACLY and the Warburg effect in a feed-back loop. Pending the development of specific inhibitors and tailored methods for identifying which specific metabolism is involved in the development of each tumor, ACLY could be targeted by inhibitors such as hydroxycitrate and bempedoic acid. The administration of citrate at high level mimics a strong inhibition of ACLY and could be tested to strengthen the effects of current therapies.

Keywords:  ACLY; AKT; acetyl-CoA; citrate; metabolism; oxaloacetate

DOI:  https://doi.org/10.1016/j.canlet.2019.12.010
Aging (Albany NY). 2019 Dec 08. 11

Histone proteomics reveals novel post-translational modifications in breast cancer.

Angela Mena Perri, Valter Agosti, Erika Olivo, Antonio Concolino, MariaTeresa De Angelis, Laura Tammè, Claudia Vincenza Fiumara, Giovanni Cuda, Domenica Scumaci.

  Histones and their variants are subjected to several post-translational modifications (PTMs). Histones PTMs play an important role in the regulation of gene expression and are critical for the development and progression of many types of cancer, including breast cancer. In this study, we used two-dimensional TAU/SDS electrophoresis, coupled with mass spectrometry for a comprehensive profiling of histone PTMs in breast cancer cell lines.Proteomic approach allowed us to identify 85 histone PTMs, seventeen of which are not reported in the UniProt database. Western blot analysis was performed to confirm a peculiar pattern of PTMs in the sporadic and hereditary breast cancer cell lines compared to normal cells. Overlapping mass spectrometry data with western blotting results, we identified, for the first time to our knowledge, a tyrosine phosphorylation on histone H1, which is significantly higher in breast cancer cells. Additionally, by inhibiting specific signaling paths, such as PI3K, PPARγ and FAK pathways, we established a correlation between their regulation and the presence of new histone PTMs. Our results may provide new insight on the possible implication of these modifications in breast cancer and may offer new perspectives for future clinical applications.

Keywords:  2D TAU gel; FAK; breast cancer; histone; phosphorylation

DOI:  https://doi.org/10.18632/aging.102577
Adv Exp Med Biol. 2019 ;1188 239-249

Utility of Reverse-Phase Protein Array for Refining Precision Oncology.

Mari Masuda, Tesshi Yamada.

  Despite the early successes of targeted therapies and continuous improvements in next-generation sequencing technology over the last two decades, genomics-driven precision oncology has helped only a minority of cancer patients; thus treatment regimens are still not matched to the vast majority of cancer patients. It has become apparent that genomic profiling in itself is limited with respect to optimal selection of patients for targeted therapy. Proteomics-based approaches (in contrast to genomics-based and transcriptomics-based approaches) capture biological processes (e.g., diversity of protein expression patterns and post-translational modifications) directly contributing to cancer pathogenesis. This encourages incorporation of concordant proteomic analyses into the next stage of precision oncology. Reverse-phase protein array (RPPA) is well suited to pharmacodynamic analysis due to its ability to precisely map signaling status using limited amounts of clinical sample. In addition, the cost-effectiveness and rapid turnaround time of the RPPA platform offer a substantial advantage over existing molecular profiling technologies in a clinical setting. In this chapter, we begin by reviewing the current status of genomics-driven precision oncology, along with its limitations and challenges. Finally, we discuss the utility of RPPA technology as a means of improving precision oncology.

Keywords:  Biomarker; Companion diagnostics; Drug resistance; Precision oncology; Reverse-phase protein array; Tissue-agnostic biomarker

DOI:  https://doi.org/10.1007/978-981-32-9755-5_13
Trends Cancer. 2019 Dec;pii: S2405-8033(19)30219-5. [Epub ahead of print]5(12): 809-821

The Metabolic Interplay between Cancer and Other Diseases.

Anne Le, Sunag Udupa, Cissy Zhang.

  Over the past decade, knowledge of cancer metabolism has expanded exponentially and has provided several clinically relevant targets for cancer therapy. Although these current approaches have shown promise, there are very few studies showing how seemingly unrelated metabolic processes in other diseases can readily occur in cancer. Moreover, the striking metabolic overlap between cancer and other diseases such as diabetes, cardiovascular, neurological, obesity, and aging has provided key therapeutic strategies that have even begun to be translated into clinical trials. These promising results necessitate consideration of the interconnected metabolic network while studying the metabolism of cancer. This review article discusses how cancer metabolism is intertwined with systemic metabolism and how knowledge from other diseases can help to broaden therapeutic opportunities for cancer.

Keywords:  aging; cancer metabolism; cardiovascular diseases; diabetes; neurological diseases; obesity

DOI:  https://doi.org/10.1016/j.trecan.2019.10.012
Cell Death Differ. 2019 Dec 13.

Autophagy in cancer: moving from understanding mechanism to improving therapy responses in patients.

Jean M Mulcahy Levy, Andrew Thorburn.

Autophagy allows for cellular material to be delivered to lysosomes for degradation resulting in basal or stress-induced turnover of cell components that provide energy and macromolecular precursors. These activities are thought to be particularly important in cancer where both tumor-promoting and tumor-inhibiting functions of autophagy have been described. Autophagy has also been intricately linked to apoptosis and programmed cell death, and understanding these interactions is becoming increasingly important in improving cancer therapy and patient outcomes. In this review, we consider how recent discoveries about how autophagy manipulation elicits its effects on cancer cell behavior can be leveraged to improve therapeutic responses.

DOI: https://doi.org/10.1038/s41418-019-0474-7