bims-pideca Biomed News
on Class IA PI3K signalling in development and cancer
Issue of 2020–12–13
thirteen papers selected by
Ralitsa Radostinova Madsen, University College London
  1. AKT1 E17K inhibits cancer cell migration by abrogating β-catenin signaling.
  2. Phosphorylation and Driver Mutations in PI3Kα and PTEN Autoinhibition.
  3. Mechanisms by which adiponectin reverses high fat diet-induced insulin resistance in mice.
  4. Blood and lymphatic systems are segregated by the FLCN tumor suppressor.
  5. Cell non-autonomous regulation of health and longevity.
  6. Lack of PTEN in osteocytes increases circulating phosphate concentrations by decreasing intact fibroblast growth factor 23 levels.
  7. Neddylation of PTEN regulates its nuclear import and promotes tumor development.
  8. Targeted inhibition of cooperative mutation- and therapy-induced AKT activation in AML effectively enhances response to chemotherapy.
  9. Targeting phosphatidylinositol 3-kinase gamma (PI3Kγ): Discovery and development of its selective inhibitors.
  10. Autocrine insulin pathway signaling regulates actin dynamics in cell wound repair.
  11. mTOR inhibition acts as an unexpected checkpoint in p53-mediated tumor suppression.
  12. Arterialization requires the timely suppression of cell growth.
  13. Integrating Quantitative Assays with Biologically Based Mathematical Modeling for Predictive Oncology.
  1. Mol Cancer Res. 2020 Dec 10. pii: molcanres.0623.2020. [Epub ahead of print]
    AKT1 E17K inhibits cancer cell migration by abrogating β-catenin signaling.
    Sizhi Paul Gao, Amber J Kiliti, Kai Zhang, Naresh Vasani, Ninghui Mao, Emmet Jordan, Hannah C Wise, Tripti Shrestha Bhattarai, Wenhuo Hu, Madeline Dorso, James A Rodrigues, Kwanghee Kim, Aphrothiti J Hanrahan, Pedram Razavi, Brett Carver, Sarat Chandarlapaty, Jorge S Reis-Filho, Barry S Taylor, David B Solit.
      Mutational activation of the PI3 kinase/AKT pathway is among the most common pro-oncogenic events in human cancers. The clinical utility of PI3K and AKT inhibitors has, however, been modest to date. Here, we used CRISPR-mediated gene editing to study the biologic consequences of AKT1 E17K mutation by developing an AKT1 E17K-mutant isogenic system in a TP53-null background. AKT1 E17K expression under the control of its endogenous promoter enhanced cell growth and colony formation, but had a paradoxical inhibitory effect on cell migration and invasion. The mechanistic basis by which activated AKT1 inhibited cell migration and invasion was increased E-cadherin expression mediated by suppression of ZEB1 transcription via altered β-catenin subcellular localization. This phenotypic effect was AKT1-specific, as AKT2 activation had the opposite effect, a reduction in E-cadherin expression. Consistent with the opposing effects of AKT1 and AKT2 activation on E-cadherin expression, a pro-migratory effect of AKT1 activation was not observed in breast cancer cells with PTEN loss or expression of an activating PIK3CA mutation, alterations which induce the activation of both AKT isoforms. The results suggest that the use of AKT inhibitors in breast cancer patients could paradoxically accelerate metastatic progression in some genetic contexts and may explain the frequent co-selection for CDH1 mutations in AKT1 mutated breast tumors. Implications: AKT1 E17K mutation in breast cancer impairs migration/invasiveness via sequestration of β-catenin to the cell membrane leading to decreased ZEB1 transcription, resulting in increased E-cadherin expression and a reversal of epithelial-mesenchymal transition.
    DOI:  https://doi.org/10.1158/1541-7786.MCR-20-0623
  2. Mol Cancer Res. 2020 Dec 07. pii: molcanres.0818.2020. [Epub ahead of print]
    Phosphorylation and Driver Mutations in PI3Kα and PTEN Autoinhibition.
    Ruth Nussinov, Mingzhen Zhang, Chung-Jung Tsai, Hyunbum Jang.
      PI3K and PTEN are the second and third most highly mutated proteins in cancer following only p53. Their actions oppose each other. PI3K phosphorylates signaling lipid PIP2 to PIP3. PTEN dephosphorylates it back. Driver mutations in both proteins accrue PIP3. PIP3 recruits Akt and PDK1 to the membrane, promoting cell cycle progression. Here we review phosphorylation events and mutations in autoinhibition in PI3K and PTEN from the structural standpoint. Our purpose is to clarify how they control the autoinhibited states. In autoinhibition a segment or a subunit of the protein occludes its functional site. Protein-protein interfaces are often only marginally stable, making them sensitive to changes in conditions in living cells. Phosphorylation can stabilize or destabilize the interfaces. Driver mutations commonly destabilize them. In analogy to 'passenger mutations', we coin 'passenger phosphorylation' to emphasize that the presence of a phosphorylation recognition sequence logo does not necessarily imply function. Rather, it may simply reflect a statistical occurrence. In both PI3K and PTEN autoinhibiting phosphorylation events are observed in the occluding 'piece'. In PI3Kα the 'piece' is the p85α subunit. In PTEN it is the C-terminal segment. In both enzymes the stabilized interface covers the domain that attaches to the membrane. Driver mutations that trigger rotation of the occluding piece or its deletion prompt activation. To date, both enzymes lack specific, potent drugs. We discuss the implications of detailed structural and mechanistic insight into oncogenic activation and how it can advance allosteric precision oncology.
    DOI:  https://doi.org/10.1158/1541-7786.MCR-20-0818
  3. Proc Natl Acad Sci U S A. 2020 Dec 08. pii: 201922169. [Epub ahead of print]
    Mechanisms by which adiponectin reverses high fat diet-induced insulin resistance in mice.
    Xiruo Li, Dongyan Zhang, Daniel F Vatner, Leigh Goedeke, Sandro M Hirabara, Ye Zhang, Rachel J Perry, Gerald I Shulman.
      Adiponectin has emerged as a potential therapy for type 2 diabetes mellitus, but the molecular mechanism by which adiponectin reverses insulin resistance remains unclear. Two weeks of globular adiponectin (gAcrp30) treatment reduced fasting plasma glucose, triglyceride (TAG), and insulin concentrations and reversed whole-body insulin resistance, which could be attributed to both improved insulin-mediated suppression of endogenous glucose production and increased insulin-stimulated glucose uptake in muscle and adipose tissues. These improvements in liver and muscle sensitivity were associated with ∼50% reductions in liver and muscle TAG and plasma membrane (PM)-associated diacylglycerol (DAG) content and occurred independent of reductions in total ceramide content. Reductions of PM DAG content in liver and skeletal muscle were associated with reduced PKCε translocation in liver and reduced PKCθ and PKCε translocation in skeletal muscle resulting in increased insulin-stimulated insulin receptor tyrosine1162 phosphorylation, IRS-1/IRS-2-associated PI3-kinase activity, and Akt-serine phosphorylation. Both gAcrp30 and full-length adiponectin (Acrp30) treatment increased eNOS/AMPK activation in muscle and muscle fatty acid oxidation. gAcrp30 and Acrp30 infusions also increased TAG uptake in epididymal white adipose tissue (eWAT), which could be attributed to increased lipoprotein lipase (LPL) activity. These data suggest that adiponectin and adiponectin-related molecules reverse lipid-induced liver and muscle insulin resistance by reducing ectopic lipid storage in these organs, resulting in decreased plasma membrane sn-1,2-DAG-induced nPKC activity and increased insulin signaling. Adiponectin mediates these effects by both promoting the storage of TAG in eWAT likely through stimulation of LPL as well as by stimulation of AMPK in muscle resulting in increased muscle fat oxidation.
    Keywords:  adiponectin; ceramides; diacylglycerol; lipoprotein lipase; protein kinase C
    DOI:  https://doi.org/10.1073/pnas.1922169117
  4. Nat Commun. 2020 12 09. 11(1): 6314
    Blood and lymphatic systems are segregated by the FLCN tumor suppressor.
    Ikue Tai-Nagara, Yukiko Hasumi, Dai Kusumoto, Hisashi Hasumi, Keisuke Okabe, Tomofumi Ando, Fumio Matsuzaki, Fumiko Itoh, Hideyuki Saya, Chang Liu, Wenling Li, Yoh-Suke Mukouyama, W Marston Linehan, Xinyi Liu, Masanori Hirashima, Yutaka Suzuki, Shintaro Funasaki, Yorifumi Satou, Mitsuko Furuya, Masaya Baba, Yoshiaki Kubota.
      Blood and lymphatic vessels structurally bear a strong resemblance but never share a lumen, thus maintaining their distinct functions. Although lymphatic vessels initially arise from embryonic veins, the molecular mechanism that maintains separation of these two systems has not been elucidated. Here, we show that genetic deficiency of Folliculin, a tumor suppressor, leads to misconnection of blood and lymphatic vessels in mice and humans. Absence of Folliculin results in the appearance of lymphatic-biased venous endothelial cells caused by ectopic expression of Prox1, a master transcription factor for lymphatic specification. Mechanistically, this phenotype is ascribed to nuclear translocation of the basic helix-loop-helix transcription factor Transcription Factor E3 (TFE3), binding to a regulatory element of Prox1, thereby enhancing its venous expression. Overall, these data demonstrate that Folliculin acts as a gatekeeper that maintains separation of blood and lymphatic vessels by limiting the plasticity of committed endothelial cells.
    DOI:  https://doi.org/10.1038/s41467-020-20156-6
  5. Elife. 2020 Dec 10. pii: e62659. [Epub ahead of print]9
    Cell non-autonomous regulation of health and longevity.
    Hillary A Miller, Elizabeth S Dean, Scott D Pletcher, Scott F Leiser.
      As the demographics of the modern world skew older, understanding and mitigating the effects of aging is increasingly important within biomedical research. Recent studies in model organisms demonstrate that the aging process is frequently modified by an organism's ability to perceive and respond to changes in its environment. Many well-studied pathways that influence aging involve sensory cells, frequently neurons, that signal to peripheral tissues and promote survival during the presence of stress. Importantly, this activation of stress response pathways is often sufficient to improve health and longevity even in the absence of stress. Here, we review the current landscape of research highlighting the importance of cell non-autonomous signaling in modulating aging from C. elegans to mammals. We also discuss emerging concepts including retrograde signaling, approaches to mapping these networks, and development of potential therapeutics.
    Keywords:  C. elegans; D. melanogaster; aging; genetics; genomics; healthspan; insulin signaling; neuroscience; sensory perception
    DOI:  https://doi.org/10.7554/eLife.62659
  6. Sci Rep. 2020 Dec 09. 10(1): 21501
    Lack of PTEN in osteocytes increases circulating phosphate concentrations by decreasing intact fibroblast growth factor 23 levels.
    Masanobu Kawai, Saori Kinoshita, Keiichi Ozono, Toshimi Michigami.
      Fibroblast growth factor 23 (FGF23) has been centric to the regulation of phosphate (Pi) metabolism; however, the regulatory network of FGF23 in osteocytes has not yet been defined in detail. We herein investigated the role of PTEN (phosphatase and tensin homolog deleted from chromosome 10) in this regulation. We created mice lacking PTEN expression mainly in osteocytes by crossing Pten-flox mice with Dmp1-Cre mice. The lack of PTEN in the osteocytes of these mice was associated with decreased skeletal and serum intact FGF23 levels, which, in turn, resulted in reductions of urinary Pi excretion and elevations of serum Pi levels. Mechanistically, the knockdown of PTEN expression in osteoblastic UMR106 cells activated the AKT/mTORC1 (mechanistic target of rapamycin complex 1) pathway and this was associated with reductions in Fgf23 expression. Furthermore, the suppression of Fgf23 expression by PTEN knockdown or insulin simulation in UMR106 cells was partially restored by the treatment with the mTORC1 inhibitor, rapamycin. These results suggest that FGF23 expression in osteoblastic cells is in part regulated through the AKT/mTORC1 pathway and provide new insights into our understanding of the regulatory network of Pi metabolism.
    DOI:  https://doi.org/10.1038/s41598-020-78692-6
  7. Cell Res. 2020 Dec 09.
    Neddylation of PTEN regulates its nuclear import and promotes tumor development.
    Ping Xie, Zhiqiang Peng, Yujiao Chen, Hongchang Li, Mengge Du, Yawen Tan, Xin Zhang, Zhe Lu, Chun-Ping Cui, Cui Hua Liu, Fuchu He, Lingqiang Zhang.
      PTEN tumor suppressor opposes the PI3K/Akt signaling pathway in the cytoplasm and maintains chromosomal integrity in the nucleus. Nucleus-cytoplasm shuttling of PTEN is regulated by ubiquitylation, SUMOylation and phosphorylation, and nuclear PTEN has been proposed to exhibit tumor-suppressive functions. Here we show that PTEN is conjugated by Nedd8 under high glucose conditions, which induces PTEN nuclear import without effects on PTEN stability. PTEN neddylation is promoted by the XIAP ligase and removed by the NEDP1 deneddylase. We identify Lys197 and Lys402 as major neddylation sites on PTEN. Neddylated PTEN accumulates predominantly in the nucleus and promotes rather than suppresses cell proliferation and metabolism. The nuclear neddylated PTEN dephosphorylates the fatty acid synthase (FASN) protein, inhibits the TRIM21-mediated ubiquitylation and degradation of FASN, and then promotes de novo fatty acid synthesis. In human breast cancer tissues, neddylated PTEN correlates with tumor progression and poor prognosis. Therefore, we demonstrate a previously unidentified pool of nuclear PTEN in the Nedd8-conjugated form and an unexpected tumor-promoting role of neddylated PTEN.
    DOI:  https://doi.org/10.1038/s41422-020-00443-z
  8. Leukemia. 2020 Dec 09.
    Targeted inhibition of cooperative mutation- and therapy-induced AKT activation in AML effectively enhances response to chemotherapy.
    Montserrat Estruch, Kristian Reckzeh, Camilla Vittori, Anders Centio, Mina Ali, Sophia Engelhard, Ling Zhao, Kyoung Jae Won, Paul Liu, Bo Torben Porse, Kim Theilgaard-Mönch.
      Most AML patients exhibit mutational activation of the PI3K/AKT signaling pathway, which promotes downstream effects including growth, survival, DNA repair, and resistance to chemotherapy. Herein we demonstrate that the inv(16)/KITD816Y AML mouse model exhibits constitutive activation of PI3K/AKT signaling, which was enhanced by chemotherapy-induced DNA damage through DNA-PK-dependent AKT phosphorylation. Strikingly, inhibitors of either PI3K or DNA-PK markedly reduced chemotherapy-induced AKT phosphorylation and signaling leading to increased DNA damage and apoptosis of inv(16)/KITD816Y AML cells in response to chemotherapy. Consistently, combinations of chemotherapy and PI3K or DNA-PK inhibitors synergistically inhibited growth and survival of clonogenic AML cells without substantially inhibiting normal clonogenic bone marrow cells. Moreover, treatment of inv(16)/KITD816Y AML mice with combinations of chemotherapy and PI3K or DNA-PK inhibitors significantly prolonged survival compared to untreated/single-treated mice. Mechanistically, our findings implicate that constitutive activation of PI3K/AKT signaling driven by mutant KIT, and potentially other mutational activators such as FLT3 and RAS, cooperates with chemotherapy-induced DNA-PK-dependent activation of AKT to promote survival, DNA repair, and chemotherapy resistance in AML. Hence, our study provides a rationale to select AML patients exhibiting constitutive PI3K/AKT activation for simultaneous treatment with chemotherapy and inhibitors of DNA-PK and PI3K to improve chemotherapy response and clinical outcome.
    DOI:  https://doi.org/10.1038/s41375-020-01094-0
  9. Med Res Rev. 2020 Dec 10.
    Targeting phosphatidylinositol 3-kinase gamma (PI3Kγ): Discovery and development of its selective inhibitors.
    Jingyu Zhu, Kan Li, Li Yu, Yun Chen, Yanfei Cai, Jian Jin, Tingjun Hou.
      Phosphatidylinositol 3-kinase gamma (PI3Kγ) has been regarded as a promising drug target for the treatment of advanced solid tumors, leukemia, lymphoma, and inflammatory and autoimmune diseases. However, the high level of structural conservation among the members of the PI3K family and the diverse physiological roles of Class I PI3K isoforms (α, β, δ, and γ) highlight the importance of isoform selectivity in the development of PI3Kγ inhibitors. In this review, we provide an overview of the structural features of PI3Kγ that influence γ-isoform selectivity and discuss the structure-selectivity-activity relationship of existing clinical PI3Kγ inhibitors. Additionally, we summarize the experimental and computational techniques utilized to identify PI3Kγ inhibitors. The insights gained so far could be used to overcome the main challenges in development and accelerate the discovery of PI3Kγ-selective inhibitors.
    Keywords:  IPI-549; PI3Kγ; isoform selective inhibitors; molecular modeling; phosphoinositide 3-kinase gamma; structure-activity relationship (SAR)
    DOI:  https://doi.org/10.1002/med.21770
  10. PLoS Genet. 2020 Dec 11. 16(12): e1009186
    Autocrine insulin pathway signaling regulates actin dynamics in cell wound repair.
    Mitsutoshi Nakamura, Jeffrey M Verboon, Tessa E Allen, Maria Teresa Abreu-Blanco, Raymond Liu, Andrew N M Dominguez, Jeffrey J Delrow, Susan M Parkhurst.
      Cells are exposed to frequent mechanical and/or chemical stressors that can compromise the integrity of the plasma membrane and underlying cortical cytoskeleton. The molecular mechanisms driving the immediate repair response launched to restore the cell cortex and circumvent cell death are largely unknown. Using microarrays and drug-inhibition studies to assess gene expression, we find that initiation of cell wound repair in the Drosophila model is dependent on translation, whereas transcription is required for subsequent steps. We identified 253 genes whose expression is up-regulated (80) or down-regulated (173) in response to laser wounding. A subset of these genes were validated using RNAi knockdowns and exhibit aberrant actomyosin ring assembly and/or actin remodeling defects. Strikingly, we find that the canonical insulin signaling pathway controls actin dynamics through the actin regulators Girdin and Chickadee (profilin), and its disruption leads to abnormal wound repair. Our results provide new insight for understanding how cell wound repair proceeds in healthy individuals and those with diseases involving wound healing deficiencies.
    DOI:  https://doi.org/10.1371/journal.pgen.1009186
  11. Genes Dev. 2020 Dec 10.
    mTOR inhibition acts as an unexpected checkpoint in p53-mediated tumor suppression.
    Ning Kon, Yang Ou, Shang-Jui Wang, Huan Li, Anil K Rustgi, Wei Gu.
      Here, we showed that the acetylation-defective p53-4KR mice, lacking the ability of cell cycle arrest, senescence, apoptosis, and ferroptosis, were tumor prone but failed to develop early-onset tumors. By identifying a novel p53 acetylation site at lysine K136, we found that simultaneous mutations at all five acetylation sites (p53-5KR) diminished its remaining tumor suppression function. Moreover, the embryonic lethality caused by the deficiency of mdm2 was fully rescued in the background of p535KR/5KR , but not p534KR/4KR background. p53-4KR retained the ability to suppress mTOR function but this activity was abolished in p53-5KR cells. Notably, the early-onset tumor formation observed in p535KR/5KR and p53-null mice was suppressed upon the treatment of the mTOR inhibitor. These results suggest that p53-mediated mTOR regulation plays an important role in both embryonic development and tumor suppression, independent of cell cycle arrest, senescence, apoptosis, and ferroptosis.
    Keywords:  DDIT4/REDD1; Mdm2; SESN2; acetylation; mTOR; p53; transcriptional regulation; tumor suppression
    DOI:  https://doi.org/10.1101/gad.340919.120
  12. Nature. 2020 Dec 09.
    Arterialization requires the timely suppression of cell growth.
    Wen Luo, Irene Garcia-Gonzalez, Macarena Fernández-Chacón, Verónica Casquero-Garcia, Maria S Sanchez-Muñoz, Severin Mühleder, Lourdes Garcia-Ortega, Jorge Andrade, Michael Potente, Rui Benedito.
      The formation of arteries is thought to occur by the induction of a highly conserved arterial genetic programme in a subset of vessels that will later experience an increase in oxygenated blood flow1,2. The initial steps of arterial specification require both the VEGF and Notch signalling pathways3-5. Here, we combine inducible genetic mosaics and transcriptomics to modulate and define the function of these signalling pathways in cell proliferation, arteriovenous differentiation and mobilization. We show that endothelial cells with high levels of VEGF or Notch signalling are intrinsically biased to mobilize and form arteries; however, they are not genetically pre-determined, and can also form veins. Mechanistically, we found that increased levels of VEGF and Notch signalling in pre-arterial capillaries suppresses MYC-dependent metabolic and cell-cycle activities, and promotes the incorporation of endothelial cells into arteries. Mosaic lineage-tracing studies showed that endothelial cells that lack the Notch-RBPJ transcriptional activator complex rarely form arteries; however, these cells regained the ability to form arteries when the function of MYC was suppressed. Thus, the development of arteries does not require the direct induction of a Notch-dependent arterial differentiation programme, but instead depends on the timely suppression of endothelial cell-cycle progression and metabolism, a process that precedes arterial mobilization and complete differentiation.
    DOI:  https://doi.org/10.1038/s41586-020-3018-x
  13. iScience. 2020 Dec 18. 23(12): 101807
    Integrating Quantitative Assays with Biologically Based Mathematical Modeling for Predictive Oncology.
    Anum S Kazerouni, Manasa Gadde, Andrea Gardner, David A Hormuth, Angela M Jarrett, Kaitlyn E Johnson, Ernesto A B F Lima, Guillermo Lorenzo, Caleb Phillips, Amy Brock, Thomas E Yankeelov.
      We provide an overview on the use of biological assays to calibrate and initialize mechanism-based models of cancer phenomena. Although artificial intelligence methods currently dominate the landscape in computational oncology, mathematical models that seek to explicitly incorporate biological mechanisms into their formalism are of increasing interest. These models can guide experimental design and provide insights into the underlying mechanisms of cancer progression. Historically, these models have included a myriad of parameters that have been difficult to quantify in biologically relevant systems, limiting their practical insights. Recently, however, there has been much interest calibrating biologically based models with the quantitative measurements available from (for example) RNA sequencing, time-resolved microscopy, and in vivo imaging. In this contribution, we summarize how a variety of experimental methods quantify tumor characteristics from the molecular to tissue scales and describe how such data can be directly integrated with mechanism-based models to improve predictions of tumor growth and treatment response.
    Keywords:  Bioengineering; Cancer; In Silico Biology; Systems Biology
    DOI:  https://doi.org/10.1016/j.isci.2020.101807
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