extracellular vesicles in vascular calcification

This Month April 2016 A summary of the most recent articles in The Journal of Clinical Investigation and JCI Insight

Also in this issue: NOTCH mediates bone fracture repair 7 Schwann cells promote cancer cell invasion 11 Review Series: Extracellular vesicles edited by Laurence Zitvogel 13 JCI Insight 14

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Extracellular vesicles in vascular calcification p. 1

On the JCI cover

This Month April 2016

For the JCI and JCI Insight Editor Howard A. Rockman Executive Editor Sarah C. Jackson Science Editors Jillian Hurst, Corinne Williams Assistant Science Editor Elyse Dankoski For the JCI Deputy Editors Garnett Kelsoe, Bryan L. Roth Associate Editors Soman N. Abraham, Vann Bennett, Gerard C. Blobe, Kathleen M. Caron, Marc G. Caron, John P. Chute, Thomas M. Coffman, Anna Mae Diehl, Ronald J. Falk, Michael B. Kastan, Daniel P. Kelly, Mary E. Klotman, Rodger A. Liddle, Nigel Mackman, Larry G. Moss, Deborah M. Muoio, Christopher B. Newgard, Paul W. Noble, Geoffrey S. Pitt, Jeffrey C. Rathmell, W. Kimryn Rathmell, Jonathan S. Serody, Norman Sharpless, Thomas Weber, Yiping Yang Clinical Medicine Associate Editors Michael A. Morse, Andrew J. Muir, Scott M. Palmer, Mark A. Stacy Asia Editor David M. Virshup Chair, Executive Council Robert J. Lefkowitz Biostatisticians Cynthia Coffman, Barry Moser, Maren Olsen Bioethicist Arthur L. Caplan Editor at Large Ushma S. Neill ISSN 2324-7703 (print) ISSN 2325-4556 (online) The American Society for Clinical Investigation holds the rights to and publishes the Journal of Clinical Investigation and JCI Insight. The opinions expressed herein are solely those of the authors and are not necessarily endorsed by the ASCI.

Extracellular vesicle–mediated vascular calcification is regulated by sortilin With age, many individuals will develop vascular calcification, which reduces arterial elasticity and is associated with increased risk of cardiovascular complications. Previous studies have demonstrated that smooth muscle cells express several genes involved in bone formation, resulting in the deposition of a calcium-mineralized matrix. Smooth muscle cells also release extracellular vesicles that contribute to calcification, but the mechanisms governing this process have not been clear. In this issue of the JCI, Elena Aikawa and colleagues report that the multiligand sorting receptor sortilin regulates loading of tissue nonspecific alkaline phosphatase (TNAP) into extracellular vesicles, which promotes calcification. They found that sortilin levels are elevated in calcified human atheroma and in a mouse model of calcified arteries. Further, mice with genetic loss of sortilin had decreased arterial calcification. Mass spectrometric analysis of sortilin-associated proteins revealed an interaction with TNAP, and subsequent studies showed that sortilin promotes TNAP activity in extracellular vesicles and microcalcification in cultured smooth muscle cells. Additionally, the research team found that smooth muscle cell calcification required Rab11, which regulates vesicle exocytosis, as well as phosphorylation of the C-termainal tail of sortilin. These findings collectively uncover a critical role for sortilin in vascular calcification induced by extracellular vesicles. The accompanying image is a density-dependent color scanning electron micrograph of a calcified human carotid artery atherosclerotic plaque. Dense, calcified areas are shown in orange, and less dense components of the plaque are shown in green. Image credit: Sergio Bertazzo. Sortilin mediates vascular calcification via its recruitment into extracellular vesicles Claudia Goettsch, Joshua D. Hutcheson, Masanori Aikawa, Hiroshi Iwata, Tan Pham, Anders Nykjaer, Mads Kjolby, Maximillian Rogers, Thomas Michel, Manabu Shibasaki, Sumihiko Hagita, Rafael Kramann, Daniel J. Rader, Peter Libby, Sasha A. Singh, and Elena Aikawa http://jci.me/80851

For the full JCI online: jci.me/126/4 For JCI Insight: jci.me/insight/1/3

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april 2016

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Research articles in the current issue of the JCI AIDS/HIV Macrophages sustain HIV replication in vivo independently of T cells Jenna B. Honeycutt, Angela Wahl, Caroline Baker, Rae Ann Spagnuolo, John Foster, Oksana Zakharova, Stephen Wietgrefe, Carolina Caro-Vegas, Victoria Madden, Garrett Sharpe, Ashley T. Haase, Joseph J. Eron, and J. Victor Garcia http://jci.me/84456 More, p. 6

Bone biology NOTCH signaling in skeletal progenitors is critical for fracture repair Fracture union

Cuicui Wang, Jason A. Inzana, Anthony J. Mirando, Yinshi Ren, Zhaoyang Liu, Jie Shen, Regis J. O’Keefe, Hani A. Awad, and Matthew J. Hilton http://jci.me/80672 More, p. 7

Endocrinology PI3-kinase mutation linked to insulin and growth factor resistance in vivo Jonathon N. Winnay, Marie H. Solheim, Ercument Dirice, Masaji Sakaguchi, Hye-Lim Noh, Hee Joon Kang, Hirokazu Takahashi, Kishan K. Chudasama, Jason K. Kim, Anders Molven, C. Ronald Kahn, and Pål R. Njølstad http://jci.me/84005

Genetics Modulation of LMNA splicing as a strategy to treat prelamin A diseases Aortic progerin

John M. Lee, Chika Nobumori, Yiping Tu, Catherine Choi, Shao H. Yang, Hea-Jin Jung, Timothy A. Vickers, Frank Rigo, C. Frank Bennett, Stephen G. Young, and Loren G. Fong http://jci.me/85908 With related Commentary by Elizabeth M. McNally and Eugene J. Wyatt More, p. 8

Hematology Renal epithelium regulates erythropoiesis via HIF-dependent suppression of erythropoietin Navid M. Farsijani, Qingdu Liu, Hanako Kobayashi, Olena Davidoff, Feng Sha, Joachim Fandrey, T. Alp Ikizler, Paul M. O’Connor, and Volker H. Haase http://jci.me/74997 Hepatic erythropoietin

RNA-binding protein IGF2BP3 targeting of oncogenic transcripts promotes hematopoietic progenitor proliferation

Jayanth Kumar Palanichamy, Tiffany M. Tran, Jonathan M. Howard, Jorge R. Contreras, Thilini R. Fernando, Timothy Sterne-Weiler, Sol Katzman, Masoud Toloue, Weihong Yan, Giuseppe Basso, Martina Pigazzi, Jeremy R. Sanford, and Dinesh S. Rao http://jci.me/80046 2

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Research articles in the current issue of the JCI

Immunology Disabled homolog 2 controls macrophage phenotypic polarization and adipose tissue inflammation Samantha E. Adamson, Rachael Griffith, Radim Moravec, Subramanian Senthivinayagam, Garren Montgomery, Wenshu Chen, Jenny Han, Poonam R. Sharma, Garrett R. Mullins, Stacey A. Gorski, Jonathan A. Cooper, Alexandra Kadl, Kyle Enfield, Thomas J. Braciale, Thurl E. Harris, and Norbert Leitinger http://jci.me/79590 More, p. 12

Estrogen-mediated downregulation of AIRE influences sexual dimorphism in autoimmune diseases Nadine Dragin, Jacky Bismuth, Géraldine Cizeron-Clairac, Maria Grazia Biferi, Claire Berthault, Alain Serraf, Rémi Nottin, David Klatzmann, Ana Cumano, Martine Barkats, Rozen Le Panse, and Sonia Berrih-Aknin http://jci.me/81894 With related Commentary by Pearl Bakhru and Maureen A. Su More, p. 12

H7N9 influenza virus neutralizing antibodies that possess few somatic mutations Natalie J. Thornburg, Heng Zhang, Sandhya Bangaru, Gopal Sapparapu, Nurgun Kose, Rebecca M. Lampley, Robin G. Bombardi, Yingchun Yu, Stephen Graham, Andre Branchizio, Sandra M. Yoder, Michael T. Rock, C. Buddy Creech, Kathryn M. Edwards, David Lee, Sheng Li, Ian A. Wilson, Adolfo García-Sastre, Randy A. Albrecht, and James E. Crowe Jr. http://jci.me/85317

Muscle biology UTX demethylase activity is required for satellite cell–mediated muscle regeneration UTX inhibition in muscle

Hervé Faralli, Chaochen Wang, Kiran Nakka, Aissa Benyoucef, Soji Sebastian, Lenan Zhuang, Alphonse Chu, Carmen G. Palii, Chengyu Liu, Brendan Camellato, Marjorie Brand, Kai Ge, and F. Jeffrey Dilworth http://jci.me/83239 With related Commentary by Ling Liu and Thomas A. Rando More, p. 8

Neuroscience Astrocytes are central in the pathomechanisms of vanishing white matter VWM glial pathology

Stephanie Dooves, Marianna Bugiani, Nienke L. Postma, Emiel Polder, Niels Land, Stephen T. Horan, Anne-Lieke F. van Deijk, Aleid van de Kreeke, Gerbren Jacobs, Caroline Vuong, Jan Klooster, Maarten Kamermans, Joke Wortel, Maarten Loos, Lisanne E. Wisse, Gert C. Scheper, Truus E.M. Abbink, Vivi M. Heine, and Marjo S. van der Knaap http://jci.me/83908 More, p. 7

Dose-escalation study of octanoic acid in patients with essential tremor Bernhard Voller, Emily Lines, Gayle McCrossin, Sule Tinaz, Codrin Lungu, George Grimes, Judith Starling, Gopal Potti, Peter Buchwald, Dietrich Haubenberger, and Mark Hallett http://jci.me/83621

Oncology LNK/SH2B3 regulates IL-7 receptor signaling in normal and malignant B-progenitors Tp53–/–Lnk–/– B cell leukemia

Ying Cheng, Kudakwashe Chikwava, Chao Wu, Haibing Zhang, Anchit Bhagat, Dehua Pei, John K. Choi, and Wei Tong http://jci.me/81468

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Oncology Loss of gastrokine-2 drives premalignant gastric inflammation and tumor progression

Gastrokine-2–deficient epithelium

Trevelyan R. Menheniott, Louise O’Connor, Yok Teng Chionh, Jan Däbritz, Michelle Scurr, Benjamin N. Rollo, Garrett Z. Ng, Shelley Jacobs, Angelique Catubig, Bayzar Kurklu, Stephen Mercer, Toshinari Minamoto, David E. Ong, Richard L. Ferrero, James G. Fox, Timothy C. Wang, Philip Sutton, Louise M. Judd, and Andrew S. Giraud http://jci.me/82655 More, p. 10

MLL1 and DOT1L cooperate with meningioma-1 to induce acute myeloid leukemia Simone S. Riedel, Jessica N. Haladyna, Matthew Bezzant, Brett Stevens, Daniel A. Pollyea, Amit U. Sinha, Scott A. Armstrong, Qi Wei, Roy M. Pollock, Scott R. Daigle, Craig T. Jordan, Patricia Ernst, Tobias Neff, and Kathrin M. Bernt http://jci.me/80825 More, p. 10

Involvement of activation-induced cytidine deaminase in skin cancer development Taichiro Nonaka, Yoshinobu Toda, Hiroshi Hiai, Munehiro Uemura, Motonobu Nakamura, Norio Yamamoto, Ryo Asato, Yukari Hattori, Kazuhisa Bessho, Nagahiro Minato, and Kazuo Kinoshita http://jci.me/81522 Skin carcinogenesis

Targeting prion-like protein doppel selectively suppresses tumor angiogenesis

Taslim A. Al-Hilal, Seung Woo Chung, Jeong Uk Choi, Farzana Alam, Jooho Park, Seong Who Kim, Sang Yoon Kim, Fakhrul Ahsan, In-San Kim, and Youngro Byun http://jci.me/83427 More, p. 11

Schwann cells induce cancer cell dispersion and invasion Sylvie Deborde, Tatiana Omelchenko, Anna Lyubchik, Yi Zhou, Shizhi He, William F. McNamara, Natalya Chernichenko, Sei-Young Lee, Fernando Barajas, Chun-Hao Chen, Richard L. Bakst, Efsevia Vakiani, Shuangba He, Alan Hall, and Richard J. Wong http://jci.me/82658 Schwann cell NCAM1

With related Commentary by Salma H. Azam and Chad V. Pecot More, p. 11

Endogenous transmembrane protein UT2 inhibits pSTAT3 and suppresses hematological malignancy Dongjun Lee, Ying-Hua Wang, Demetrios Kalaitzidis, Janani Ramachandran, Homare Eda, David B. Sykes, Noopur Raje, and David T. Scadden http://jci.me/84620

Thymic stromal lymphopoietin blocks early stages of breast carcinogenesis Shadmehr Demehri, Trevor J. Cunningham, Sindhu Manivasagam, Kenneth H. Ngo, Sara Moradi Tuchayi, Rasika Reddy, Melissa A. Meyers, David G. DeNardo, and Wayne M. Yokoyama http://jci.me/83724 TSLP-induced breast cancer

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Pulmonology Macrophage-epithelial paracrine crosstalk inhibits lung edema clearance during influenza infection

Influenza-infected lung

Christin Peteranderl, Luisa Morales-Nebreda, Balachandar Selvakumar, Emilia Lecuona, István Vadász, Rory E. Morty, Carole Schmoldt, Julia Bespalowa, Thorsten Wolff, Stephan Pleschka, Konstantin Mayer, Stefan Gattenloehner, Ludger Fink, Juergen Lohmeyer, Werner Seeger, Jacob I. Sznajder, Gökhan M. Mutlu, G.R. Scott Budinger, and Susanne Herold http://jci.me/83931 With related Commentary by Rena Brauer and Peter Chen More, p. 9

Transplantation Metabolic reprogramming of alloantigen-activated T cells after hematopoietic cell transplantation Hung D. Nguyen, Shilpak Chatterjee, Kelley M.K. Haarberg, Yongxia Wu, David Bastian, Jessica Heinrichs, Jianing Fu, Anusara Daenthanasanmak, Steven Schutt, Sharad Shrestha, Chen Liu, Honglin Wang, Hongbo Chi, Shikhar Mehrotra, and Xue-Zhong Yu http://jci.me/82587 More, p. 6

Alloantigen-specific regulatory T cells generated with a chimeric antigen receptor Katherine G. MacDonald, Romy E. Hoeppli, Qing Huang, Jana Gillies, Dan S. Luciani, Paul C. Orban, Raewyn Broady, and Megan K. Levings http://jci.me/82771 With related Commentary by Matthias Edinger More, p. 6

Vascular biology Sortilin mediates vascular calcification via its recruitment into extracellular vesicles Calcified smooth muscle

Claudia Goettsch, Joshua D. Hutcheson, Masanori Aikawa, Hiroshi Iwata, Tan Pham, Anders Nykjaer, Mads Kjolby, Maximillian Rogers, Thomas Michel, Manabu Shibasaki, Sumihiko Hagita, Rafael Kramann, Daniel J. Rader, Peter Libby, Sasha A. Singh, and Elena Aikawa http://jci.me/80851 More, p. 1

CCR7 and IRF4-dependent dendritic cells regulate lymphatic collecting vessel permeability Stoyan Ivanov, Joshua P. Scallan, Ki-Wook Kim, Kathrin Werth, Michael W. Johnson, Brian T. Saunders, Peter L. Wang, Emma L. Kuan, Adam C. Straub, Melissa Ouhachi, Erica G. Weinstein, Jesse W. Williams, Carlos Briseño, Marco Colonna, Brant E. Isakson, Emmanuel L. Gautier, Reinhold Förster, Michael J. Davis, Bernd H. Zinselmeyer, and Gwendalyn J. Randolph http://jci.me/84518

Matricellular protein CCN3 mitigates abdominal aortic aneurysm

Matrix metalloproteinase activity

Chao Zhang, Dustin van der Voort, Hong Shi, Rongli Zhang, Yulan Qing, Shuichi Hiraoka, Minoru Takemoto, Koutaro Yokote, Joseph V. Moxon, Paul Norman, Laure Rittié, Helena Kuivaniemi, G. Brandon Atkins, Stanton L. Gerson, Guo-Ping Shi, Jonathan Golledge, Nianguo Dong, Bernard Perbal, Domenick A. Prosdocimo, and Zhiyong Lin http://jci.me/82337

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JCI research | Editor’s picks

transplantation

Targeting T cell metabolic alterations in graft-versushost disease Graft-versus-host disease (GVHD), which is caused by alloreactive donor T cells, limits the success of allogeneic hematopoietic stem cell transplantation (HSCT). Though metabolic alterations are known to drive changes in T cell fate and function, little is known about T cell metabolic alterations in GVHD. Hung Nguyen and colleagues examined T cell metabolic profiles using two murine models of HSCT. They found that donor T cells undergo metabolic reprogramming after transplantation, switching from tricarboxylic cycle–mediated fatty acid β oxidation (FAO) and pyruvate oxidation to aerobic glycolysis. Importantly, pharmacological or genetic inhibition of glycolysis abrogated GVHD development. These data indicate targeting glycolysis may help to control GVHD. Metabolic reprogramming of alloantigen-activated T cells after hematopoietic cell transplantation Hung D. Nguyen, Shilpak Chatterjee, Kelley M.K. Haarberg, Yongxia Wu, David Bastian, Jessica Heinrichs, Jianing Fu, Anusara Daenthanasanmak, Steven Schutt, Sharad Shrestha, Chen Liu, Honglin Wang, Hongbo Chi, Shikhar Mehrotra, and Xue-Zhong Yu http://jci.me/82587

Chimeric antigen receptor–bearing Tregs for transplantation Regulatory T cells (Tregs) are critical in preventing autoimmunity and controlling responses to alloantigens in the setting of transplantation, and Treg-based therapy is emerging as a promising strategy to prevent or treat graft-versus-host disease (GVHD) and rejection. There is emerging evidence that disease-relevant, antigen-specific Tregs are more efficacious than polyclonal Tregs; however, the generation of alloantigenspecific Tregs relies on APC-mediated expansion and requires access to both donor and recipient tissues and multiple MHC mismatches. To circumvent these hurdles, Katherine MacDonald and colleagues engineered human Tregs bearing chimeric antigen receptors (CARs) specific for HLA-A2 (A2-CAR). In a murine model of hematopoietic stem cell transplantation, A2-CAR–expressing Tregs were superior to polyclonal Tregs in preventing GVHD caused by HLA-A2+ T cells. In the accompanying Commentary, Matthias Edinger discusses how these findings indicate that administration of CAR-Tregs may be a suitable approach for generating antigen-specific Tregs. Alloantigen-specific regulatory T cells generated with a chimeric antigen receptor Katherine G. MacDonald, Romy E. Hoeppli, Qing Huang, Jana Gillies, Dan S. Luciani, Paul C. Orban, Raewyn Broady, and Megan K. Levings http://jci.me/82771 Related Commentary Driving allotolerance: CAR-expressing Tregs for tolerance induction in organ and stem cell transplantation Matthias Edinger http://jci.me/86827

aids/hiv

Human macrophages independently sustain HIV viral replication The role of macrophages in HIV infection is a topic of intense debate, and several recent studies have suggested that the presence of HIV in macrophages is the result of T cell engulfment rather than direct infection. To directly evaluate HIV replication in myeloid cells, Jenna Honeycutt and colleagues developed a humanized mouse model that is systemically reconstituted with human B cells and myeloid cells but is completely devoid of T cells (myeloid-only mice [MoM]). Using MoM, they demonstrated that myeloid cells sustained HIV replication in the absence of T cells. HIV-infected macrophages were distributed in every tissue analyzed, including the brain (see the accompanying image), where they have long been postulated to play a role in establishing HIV infection in the CNS. Moreover, infected macrophages could establish HIV infection in uninfected animals. These results demonstrate that macrophages are a target of HIV infection in vivo. Macrophages sustain HIV replication in vivo independently of T cells Jenna B. Honeycutt, Angela Wahl, Caroline Baker, Rae Ann Spagnuolo, John Foster, Oksana Zakharova, Stephen Wietgrefe, Carolina Caro-Vegas, Victoria Madden, Garrett Sharpe, Ashley T. Haase, Joseph J. Eron, and J. Victor Garcia http://jci.me/84456

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bone biology

NOTCH signaling forms a more perfect union in bones Approximately 10%–20% of fractures permanently fail to heal, a condition known as nonunion. Fracture repair appears to be achieved through both proliferation and differentiation of skeletal progenitors, but the factors governing these cells after a fracture are unknown. NOTCH signaling is required for the maintenance of bone marrow/stromal stem cells (BMSCs) during skeletal development. Cuicui Wang and colleagues examined the role of NOTCH in fracture union by examining healing of nonstabilized and stabilized fractures in mice with targeted deletion of the NOTCH signaling mediator RBPjk in BMSCs, maturing osteoblasts, and committed chondrocytes. They found that fracture nonunion only occurred in mice that lacked NOTCH signaling in BMSCs (see the accompanying image), ultimately resulting in BMSC depletion. NOTCH signaling in skeletal progenitors is critical for fracture repair Cuicui Wang, Jason A. Inzana, Anthony J. Mirando, Yinshi Ren, Zhaoyang Liu, Jie Shen, Regis J. O’Keefe, Hani A. Awad, and Matthew J. Hilton http://jci.me/80672

neuroscience

Murine models of vanishing white matter leukodystrophy Vanishing white matter (VWM) is a form of fatal leukodystrophy that is caused by mutations in eukaryotic translation initiation factor 2B (eIF2B). Stephanie Dooves, Marianna Bugiani, and colleagues developed murine models of VWM with mutations that cause severe disease in humans. Similar to humans, the mice had a highly variable phenotypic severity, and all of the strains showed impaired white matter astrocyte maturation (see the accompanying image) that was apparent prior to disease onset and increased in parallel with disease severity. The disease also affected non-forebrain astrocytes, which was not previously recognized in patients with VWM. Dooves, Bugiani, and colleagues found that VWM astrocytes secrete factors

that prevent oligodendrocyte maturation. These studies confirm that astrocytes are central to VWM pathogenesis. Astrocytes are central in the pathomechanisms of vanishing white matter Stephanie Dooves, Marianna Bugiani, Nienke L. Postma, Emiel Polder, Niels Land, Stephen T. Horan, Anne-Lieke F. van Deijk, Aleid van de Kreeke, Gerbren Jacobs, Caroline Vuong, Jan Klooster, Maarten Kamermans, Joke Wortel, Maarten Loos, Lisanne E. Wisse, Gert C. Scheper, Truus E.M. Abbink, Vivi M. Heine, and Marjo S. van der Knaap http://jci.me/83908

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muscle biology genetics

Stem cell–mediated muscle regeneration requires UTX histone demethylase activity Epigenetic-mediated cell fate specification is critical in development and stem cell–mediated tissue regeneration. Hervé Faralli and colleagues demonstrate that the histone demethylase UTX is required for satellite cell–mediated muscle regeneration. KO of Utx in female mice results in embryonic lethality; however, KO of Utx in male mice, which express a Y chromosome–encoded UTX paralog that does not have demethylase activity, survive. Faralli and colleagues generated mice with inducible, satellite cell–specific knockdown of UTX to investigate the role of UTX in muscle regeneration. Loss of UTX in satellite cells blocked myofiber regeneration (see the accompanying image). Moreover, regeneration was blocked by pharmacological inhibition of UTX demethylase activity or knock-in of an enzymatically dead mutant. Mechanistically, UTX was required for the terminal differentiation of satellite cells and the expression of a muscle gene expression program driven by histone demethylation. In the accompanying Commentary, Ling Liu and Thomas Rando discuss how these data identify a role for epigenetic regulation in myofiber regeneration. UTX demethylase activity is required for satellite cell–mediated muscle regeneration Hervé Faralli, Chaochen Wang, Kiran Nakka, Aissa Benyoucef, Soji Sebastian, Lenan Zhuang, Alphonse Chu, Carmen G. Palii, Chengyu Liu, Brendan Camellato, Marjorie Brand, Kai Ge, and F. Jeffrey Dilworth http://jci.me/83239 Related Commentary UTX in muscle regeneration — the right dose and the right time Ling Liu and Thomas A. Rando http://jci.me/86798

Shifting the balance in alternative splicing to rescue laminopathies The LMNA gene produces two alternatively spliced transcripts encoding the proteins lamin A and lamin C. Both proteins are produced in nearly equal amounts in most tissues and appear to play largely redundant roles in maintaining the nuclear lamina. Most disease-causing LMNA mutations affect both lamin A and lamin C; however, the most common LMNA mutations causing HutchinsonGilford progeria syndrome (HGPS) only affect lamin A synthesis, producing a truncated mutant form of lamin A (progerin). In this issue, John Lee and colleagues hypothesized that HGPS could be treated by shifting the balance of LMNA splicing toward lamin C to reduce the synthesis of progerin. They identified an antisense oligonucleotide (ASO) targeting exon 11 of LMNA that increased lamin C production at the expense of lamin A. Importantly, treatment of WT or HGPS mice with the ASO reduced lamin A and progerin expression, respectively. In the accompanying Commentary, Elizabeth McNally and Eugene Wyatt discuss how these findings suggest that strategies to alter LMNA splicing may be a promising treatment approach for lamin A–specific diseases. Modulation of LMNA splicing as a strategy to treat prelamin A diseases John M. Lee, Chika Nobumori, Yiping Tu, Catherine Choi, Shao H. Yang, Hea-Jin Jung, Timothy A. Vickers, Frank Rigo, C. Frank Bennett, Stephen G. Young, and Loren G. Fong http://jci.me/85908 Related Commentary Welcome to the splice age: antisense oligonucleotide–mediated exon skipping gains wider applicability Elizabeth M. McNally and Eugene J. Wyatt http://jci.me/86799

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pulmonology

Influenza-induced lung edema is mediated by macrophage/ epithelial cell crosstalk Influenza A virus (IAV) infection can cause lung injury and acute respiratory distress syndrome (ARDS) characterized by lung edema. Normally, alveolar epithelial cells (AECs) clear the excess fluid driven by the activity of the Na/K-ATPase; however, this mechanism appears to be impaired in ARDS. Christin Peteranderl and colleagues identified a paracrine communication network involving IAV-infected and uninfected AECs and alveolar macrophages, which drives decreased expression of the Na/K-ATPase, resulting in impaired alveolar fluid clearance. Mechanistic studies revealed that IAV infection induces an IFN-α/TRAIL signaling loop between AECs and macrophages that activates AMPK in AECs, triggering decreased plasma membrane expression of Na/K-ATPase. Importantly, antibody-mediated disruption of this signaling loop in mice improved the expression of Na/K-ATPase on AECs and reduced lung edema after IAV infection (see the accompanying image). In the accompanying Commentary, Rena Brauer and Peter Chen discuss how the paracrine network may be a suitable therapeutic target in lung edema.

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Macrophage-epithelial paracrine crosstalk inhibits lung edema clearance during influenza infection Christin Peteranderl, Luisa Morales-Nebreda, Balachandar Selvakumar, Emilia Lecuona, István Vadász, Rory E. Morty, Carole Schmoldt, Julia Bespalowa, Thorsten Wolff, Stephan Pleschka, Konstantin Mayer, Stefan Gattenloehner, Ludger Fink, Juergen Lohmeyer, Werner Seeger, Jacob I. Sznajder, Gökhan M. Mutlu, G.R. Scott Budinger, and Susanne Herold http://jci.me/83931 Related Commentary Influenza leaves a TRAIL to pulmonary edema Rena Brauer and Peter Chen http://jci.me/86802

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oncology

Gastrokine-2 restrains inflammation to attenuate gastric tumorigenesis Chronic inflammation due to Helicobacter pylori infection or other causes is a key risk factor in the development of gastric cancer (GC), but the mechanisms underlying this link are unclear. In this issue, Trevelyan Menheniott and colleagues analyzed gene expression in both human and murine GCs and found that loss of gastrokine-2 (GKN2) in the gastric mucosa was associated with tumor progression. Targeted deletion of Gkn2 in mice caused mucosal defects and potentiated tumorigenesis of the gastric corpus in a cytokine-driven murine GC model (gp130F/F). Conversely, overexpression of human gastrokines attenuated tumorigenesis in this model (see the accompanying image). Loss of Gkn2 in H. pylori–infected mice accelerated progression of atrophic gastritis and mucosal metaplasia that was driven by heightened innate immune responses and impaired myeloid-derived suppressor cell responses, promoting inflammation. These data establish gastrokines as critical suppressors of inflammation-driven gastric tumors. Loss of gastrokine-2 drives premalignant gastric inflammation and tumor progression Trevelyan R. Menheniott, Louise O’Connor, Yok Teng Chionh, Jan Däbritz, Michelle Scurr, Benjamin N. Rollo, Garrett Z. Ng, Shelley Jacobs, Angelique Catubig, Bayzar Kurklu, Stephen Mercer, Toshinari Minamoto, David E. Ong, Richard L. Ferrero, James G. Fox, Timothy C. Wang, Philip Sutton, Louise M. Judd, and Andrew S. Giraud http://jci.me/82655 ADVERTISEMENT

Epigenetic drivers in meningioma-1–induced acute myeloid leukemia Meningioma-1 (MN1) is frequently overexpressed in acute myeloid leukemia (AML) and is associated with poor prognosis. Forced overexpression of MN1 (MN1hi) in mice causes an aggressive myeloid leukemia that is dependent on a well-defined gene expression program, but the factors mediating this program are unknown. Simone Riedel, Jessica Haladyna, and colleagues demonstrate that two histone methyltransferases, MLL1 and DOT1L, control the gene expression program underlying MN1hi leukemia. Deletion of either Mll1 or Dot1l impaired MN1-mediated leukemogenesis and blocked expression of key leukemogenic genes. Moreover, Riedel, Haladyna, and colleagues determined that a subset of MN1hi primary patient leukemias coexpressing the MLL1 downstream target gene HOXA9 were sensitive to DOT1L inhibition, suggesting that DOT1L inhibition may be a suitable therapeutic strategy in this AML subtype. MLL1 and DOT1L cooperate with meningioma-1 to induce acute myeloid leukemia Simone S. Riedel, Jessica N. Haladyna, Matthew Bezzant, Brett Stevens, Daniel A. Pollyea, Amit U. Sinha, Scott A. Armstrong, Qi Wei, Roy M. Pollock, Scott R. Daigle, Craig T. Jordan, Patricia Ernst, Tobias Neff, and Kathrin M. Bernt http://jci.me/80825

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oncology

Targeting the prion-like protein doppel to block tumor angiogenesis Antiangiogenic therapies targeting VEGF and its receptors are used to treat a variety of solid tumors; however, these drugs target both cancerous and healthy tissues, making a tumor-specific therapy desirable. Taslim Al-Hilal and colleagues report that the prion-like protein doppel is expressed in tumor-associated vascular endothelial cells (ECs), but not in normal endothelium (see the accompanying image). In a murine s.c. tumor xenograft model, doppel overexpression in ECs enhanced tumor vascularization, while doppel inhibition reduced tumor vascularization by promoting VEGFR2 internalization and degradation in tumor-associated ECs. Administration of an orally active glycosaminoglycan,

LHbisD4, which specifically binds with doppel and concentrates over the tumor site, replicated doppel inhibition. These results indicate that doppel is a selective therapeutic target for blocking tumor angiogenesis. Targeting prion-like protein doppel selectively suppresses tumor angiogenesis Taslim A. Al-Hilal, Seung Woo Chung, Jeong Uk Choi, Farzana Alam, Jooho Park, Seong Who Kim, Sang Yoon Kim, Fakhrul Ahsan, In-San Kim, and Youngro Byun http://jci.me/83427

Cancer cells exploit neural repair pathways to enhance invasion Perineural invasion commonly occurs with a variety of malignancies and is associated with increased risk of local recurrence and poor prognosis, but the mechanisms by which nerves contribute to cancer progression are poorly understood. Using murine and human tumor specimens, Sylvie Deborde and colleagues demonstrated that a subpopulation of Schwann cells associates with cancer cells (see the accompanying image). Coculture studies revealed

that Schwann cells recruit cancer cells and induce the cancer cells to form protrusions directed toward the Schwann cells via neural cell adhesion molecule 1 (NCAM1). This interaction allows the Schwann cells to insinuate themselves between the cancer cells, dispersing them and promoting invasion. In the accompanying Commentary, Salma Azam and Chad Pecot discuss how these findings define a mechanism underlying perineural invasion.

Schwann cells induce cancer cell dispersion and invasion Sylvie Deborde, Tatiana Omelchenko, Anna Lyubchik, Yi Zhou, Shizhi He, William F. McNamara, Natalya Chernichenko, Sei-Young Lee, Fernando Barajas, Chun-Hao Chen, Richard L. Bakst, Efsevia Vakiani, Shuangba He, Alan Hall, and Richard J. Wong http://jci.me/82658 Related Commentary Cancer’s got nerve: Schwann cells drive perineural invasion Salma H. Azam and Chad V. Pecot http://jci.me/86801 jci.org/this-month

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immunology

Estrogen-induced repression of AIRE may underlie sexual dimorphism in autoimmunity Females are more susceptible to autoimmune diseases than males. Nadine Dragin and colleagues identified sex differences in thymic expression of tissue-specific antigens controlled by the autoimmune regulator (AIRE), which directly regulates the generation of autoreactive immune cells. Therefore, they tested the hypothesis that AIRE levels are linked to the sexual dimorphism of autoimmune disease susceptibility. They found that after puberty, both human and murine females expressed less AIRE in thymic epithelial cells than did males. Moreover, AIRE expression was related to sex hormones, as both male castration and estrogen administration decreased expression of AIRE in the thymus. Mechanistically, estrogen increased CpG methylation within the AIRE promoter in human thymic epithelial cells (see the accompanying image). In the related Commentary, Pearl Bakhru and Maureen Su

discuss how these studies indicate that higher estrogen levels in females induce epigenetic changes that reduce AIRE expression and increase susceptibility to autoimmune diseases. Estrogen-mediated downregulation of AIRE influences sexual dimorphism in autoimmune diseases Nadine Dragin, Jacky Bismuth, Géraldine Cizeron-Clairac, Maria Grazia Biferi, Claire Berthault, Alain Serraf, Rémi Nottin, David Klatzmann, Ana Cumano, Martine Barkats, Rozen Le Panse, and Sonia Berrih-Aknin http://jci.me/81894 Related Commentary Estrogen turns down “the AIRE” Pearl Bakhru and Maureen A. Su http://jci.me/86800

DAB2 tips the balance in macrophage polarization Macrophages change their phenotype and function in response to environmental cues generated during acute and chronic tissue injury. In order to identify regulators of macrophage phenotypic polarization, Samantha Adamson, Rachael Griffiths, and colleagues compared gene expression patterns of in vitro polarized macrophages. They found that disabled homolog 2 (DAB2) was upregulated in M2 macrophages and downregulated in M1 macrophages. Mice with myeloid-specific deletion of Dab2 showed enhanced

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inflammatory responses to LPS, with increased inflammatory gene expression and macrophage activation. Further, chronic high-fat feeding of these mice resulted in increased adipose tissue inflammation, M1 polarization of adipose tissue– resident macrophages, and insulin resistance. Mechanistically, DAB2 attenuates inflammatory NF-κB signaling through interactions with TNF receptor–associated factor 6 (TRAF6). These findings suggest that DAB2 may be a suitable therapeutic target in inflammatory disorders.

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Disabled homolog 2 controls macrophage phenotypic polarization and adipose tissue inflammation Samantha E. Adamson, Rachael Griffiths, Radim Moravec, Subramanian Senthivinayagam, Garren Montgomery, Wenshu Chen, Jenny Han, Poonam R. Sharma, Garrett R. Mullins, Stacey A. Gorski, Jonathan A. Cooper, Alexandra Kadl, Kyle Enfield, Thomas J. Braciale, Thurl E. Harris, and Norbert Leitinger http://jci.me/79590

JCI Review Series

Extracellular vesicles Series Editor: Laurence Zitvogel

Getting the message: extracellular vesicles in health and disease Cell-to-cell communication is an essential component in multicellular organisms, allowing for rapid, coordinated responses to changes within the environment. Classical signaling mediators include direct cell-cell contact as well as secreted factors, such as cytokines, metabolites, and hormones. In the past decade, extracellular vesicles (EVs), including exosomes, microvesicles, and apoptotic bodies, have emerged as important mediators of intercellular communication. EVs are doublemembrane vesicles containing cargoes of multiple proteins, lipids, and nucleic acids, which are derived from their cells of origin, and EV cargoes can change depending on the status of their originating cells. Importantly, EVs are found in all body fluids and can carry their cargoes to distant sites within the body as well as neighboring cells. Reviews in this series discuss the role of EV-mediated signaling in physiological and pathophysiological conditions, including infection, host immune responses, and cancer. Additionally, these Reviews discuss the potential clinical use of EVs as therapeutics and diagnostic biomarkers. Extracellular vesicles: masters of intercellular communication and potential clinical interventions Jonathan M. Pitt, Guido Kroemer, and Laurence Zitvogel http://jci.me/87316 Dr. Laurence Zitvogel is the Research Director at Institut National de la Santé et Recherche Médicale and a Co-Director of a Center of Clinical Investigations at Institut Gustave Roussy, Villejuif, France. She graduated in 1992 with a degree in medical oncology from the School of Medicine of the University of Paris. She began her scientific career at the University of Pittsburgh, working under Michael Lotze. Her expertise is mainly the biology of dendritic and innate effector cells, and their relevance during tumor development and cancer immunotherapy. Dr. Zitvogel currently investigates the immunogenicity of cell death with drugs of the oncological armamentarium as well as dendritic cell– derived autologous exosome vaccines. jci.org/this-month

Prostasomes as a source of diagnostic biomarkers for prostate cancer Carla Zijlstra and Willem Stoorvogel http://jci.me/81128 Extracellular vesicle isolation and characterization: toward clinical application Rong Xu, David W. Greening, Hong-Jian Zhu, Nobuhiro Takahashi, and Richard J. Simpson http://jci.me/81129 Versatile roles of extracellular vesicles in cancer Nobuyoshi Kosaka, Yusuke Yoshioka, Yu Fujita, and Takahiro Ochiya http://jci.me/81130 Regulation of chronic inflammatory and immune processes by extracellular vesicles Paul D. Robbins, Akaitz Dorronsoro, and Cori N. Booker http://jci.me/81131 Extracellular vesicles and infectious diseases: new complexity to an old story Jeffrey S. Schorey and Clifford V. Harding http://jci.me/81132 Exosomes in stroke pathogenesis and therapy Zheng Gang Zhang and Michael Chopp http://jci.me/81133 Extracellular vesicles and intercellular communication within the nervous system Valentina Zappulli, Kristina Pagh Friis, Zachary Fitzpatrick, Casey A. Maguire, and Xandra O. Breakefield http://jci.me/81134 The biology and function of exosomes in cancer Raghu Kalluri http://jci.me/81135 Exosomes and tumor-mediated immune suppression Theresa L. Whiteside http://jci.me/81136 Dendritic cell–derived exosomes for cancer therapy Jonathan M. Pitt, Fabrice André, Sebastian Amigorena, Jean-Charles Soria, Alexander Eggermont, Guido Kroemer, and Laurence Zitvogel http://jci.me/81137

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JCI Insight is a new peer-reviewed journal dedicated to biomedical research, ranging from preclinical to clinical studies.

Published March 17, 2016 Decidual Cox2 inhibition improves fetal and maternal outcomes in a preeclampsia-like mouse model Jenny L. Sones, Jeeyeon Cha, Ashley K. Woods, Amanda Bartos, Christa Y. Heyward, Heinrich E. Lob, Catherine E. Isroff, Scott D. Butler, Stephanie E. Shapiro, Sudhansu K. Dey, and Robin L. Davisson http://jci.me/75351

Trimeprazine increases IRS2 in human islets and promotes pancreatic β cell growth and function in mice

Insulin-producing cells

Alexandra Kuznetsova, Yue Yu, Jennifer Hollister-Lock, Lynn Opare-Addo, Aldo Rozzo, Marianna Sadagurski, Lisa Norquay, Jessica E. Reed, Ilham El Khattabi, Susan Bonner-Weir, Gordon C. Weir, Arun Sharma, and Morris F. White http://jci.me/80749

Maturational characteristics of HIV-specific antibodies in viremic individuals

Eric Meffre, Aaron Louie, Jason Bannock, Leo J.Y. Kim, Jason Ho, Cody C. Frear, Lela Kardava, Wei Wang, Clarisa M. Buckner, Yimeng Wang, Olivia R. Fankuchen, Kathleen R. Gittens, Tae-Wook Chun, Yuxing Li, Anthony S. Fauci, and Susan Moir http://jci.me/84610

Identification of human plasma cells with a lamprey monoclonal antibody Cuiling Yu, Yanling Liu, Justin Tze Ho Chan, Jiefei Tong, Zhihua Li, Mengyao Shi, Dariush Davani, Marion Parsons, Srijit Khan, Wei Zhan, Shuya Kyu, Eyal Grunebaum, Paolo Campisi, Evan J. Propst, David L. Jaye, Suzanne Trudel, Michael F. Moran, Mario Ostrowski, Brantley R. Herrin, F. Eun-Hyung Lee, Ignacio Sanz, Max D. Cooper, and Götz R.A. Ehrhardt http://jci.me/84738

Innate immune reconstitution with suppression of HIV-1 Eileen P. Scully, Ainsley Lockhart, Wilfredo Garcia-Beltran, Christine D. Palmer, Chelsey Musante, Eric Rosenberg, Todd M. Allen, J. Judy Chang, Ronald J. Bosch, and Marcus Altfeld http://jci.me/85433

Identifying candidate genes for 2p15p16.1 microdeletion syndrome using clinical, genomic, and functional analysis

Zebrafish rel deficiency

Hani Bagheri, Chansonette Badduke, Ying Qiao, Rita Colnaghi, Iga Abramowicz, Diana Alcantara, Christopher Dunham, Jiadi Wen, Robert S. Wildin, Malgorzata J.M. Nowaczyk, Jennifer Eichmeyer, Anna Lehman, Bruno Maranda, Sally Martell, Xianghong Shan, Suzanne M.E. Lewis, Mark O’Driscoll, Cheryl Y. Gregory-Evans, and Evica Rajcan-Separovic http://jci.me/85461

The MERTK/FLT3 inhibitor MRX-2843 overcomes resistance-conferring FLT3 mutations in acute myeloid leukemia Katherine A. Minson, Catherine C. Smith, Deborah DeRyckere, Clara Libbrecht, Alisa B. Lee-Sherick, Madeline G. Huey, Elisabeth A. Lasater, Gregory D. Kirkpatrick, Michael A. Stashko, Weihe Zhang, Craig T. Jordan, Dmitri Kireev, Xiaodong Wang, Stephen V. Frye, H. Shelton Earp, Neil P. Shah, and Douglas K. Graham http://jci.me/85630

Claudin-low bladder tumors are immune infiltrated and actively immune suppressed Jordan Kardos, Shengjie Chai, Lisle E. Mose, Sara R. Selitsky, Bhavani Krishnan, Ryoichi Saito, Michael D. Iglesia, Matthew I. Milowsky, Joel S. Parker, William Y. Kim, and Benjamin G. Vincent http://jci.me/85902

More information: http://jci.me/insinf or [email protected] 14

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