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`1 (Jan. 2010)
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`WI JUJ
`2010-01 .-.._I Q":
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`4
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`PROPERTY OF THE
`NATIONAL
`LIBRARY OF
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`The
`“ Clinical Investigation
`
`UNIV. CHICAGO EX. 2017
`
`.
`
`.. Adipose._tissue inflammatioflt‘
`
`Genome & Co. v. Univ. of Chicago
`PG R201 9-00002
`
`;
`
`

`

`Al
`ajcr Volume 120
`
`Number 1
`
`January 2010
`
`The publication of the American Socrety lor Clinical lnvealigrlllnll
`
`Editor Laurence A. Turka
`
`Deputy Editors Morris Birnbaum. Jonathan A. Epstein, Gary Korelzky
`Executive Editor Ushma S. Neill Science Editor Brooke Grindlingor
`Senior Editors
`
`Stephen G. Emerson
`
`William N. Kelley
`
`flows and Reviews Ed"
`Craig El Thompson
`
`or Karon Honev
`
`Senior European Editcir
`
`Marc Feldmann
`
`Associate Editors
`Nancy Bonini
`Michael F. Beers
`Yongwon Choi
`Ronald G. Collman
`
`Consulting Editors
`E. Dale Abel
`Domenico Accili
`Flexlord S. Ahima
`Sunil K. Ahuia
`Qais AIAAwqali
`Kari Alilalo
`Masavuki Amagai
`Moshe Arditi
`M. Amin Arnaour
`David Arlis
`Alan Allie
`Jane Aubin
`Amer Beg
`Ivor Benlamin
`Jean Bennett
`Joel S. Bennett
`Bradtord Berk
`Nina Bharrlwai
`Mina Bissotl
`Joseph V. Etonveriire
`Karin E. Brrrnleldl
`Brendan Boyce
`Thomas Bracrale
`David Allen Brenner
`Jonathan Brombern
`Vincenzo Bronte
`Michael A. Brownioe
`Hal E. Broxnicyor
`Polar Carmeliral
`Marlin Carroll
`Arturo Casadevall
`Andrew Chan
`Kvong-Mr Chang
`Harold Chapman
`lsrael Charo
`Youhai Chen
`Davrd Chereah
`Alan D. Cherrrngton
`Leonard Chess
`Angela Christiano
`Marco Colonna
`George Colsarelis
`Riccardo Dalla-Favora
`Alan D'Andrea
`Chi Van Dang
`Alan Daugherty
`
`Walik 8. El-Deiry
`Alan M. Gewirtz
`Stanley Goldlarb
`Klaus Kaestner
`Mark L. Kahn
`
`Sudhansu Day
`Betty A. Diamond
`Anna Mae Diehl
`Harry C. Diet: lll
`Stefanie Dirnmelor
`Gerald Dorn
`Michael Dustin
`Connie Eaves
`Elazer Ft. Eo‘elman
`Albert 0. Edwards
`George S. Eisenbartlr
`Leil Ellison
`Joel Elmqwsi
`Charis Eng
`.Joel D. Ernst
`Eric. Ft. F-oarori
`Harold l. Falljn'lan
`[roan Felsher
`Anthony W Ferraritr:
`Narir'ileori Ferraro
`Edward A. Irlljilel'
`‘-Jiir.nn Fisher
`lfir-irrot Fllilfiernlrl
`i’aul Frnrrritlr}
`Alan B Frov
`Joshua Friedman
`Philippe l'rnguel
`Sam {Sandy
`Donald E Garrem
`Jack Gauldre
`Allied L George Jr
`Andrew T Gowirtr
`Eli Gilhoa
`HEEI’IW Ginsberg:
`Latino Glirnollor
`Todd Golda
`Gregory J Sores.-
`Sleven Grant
`Linda Greenbaum
`Mark I. Greene
`David Haller
`Katherine A. Hailar
`Goran Hanssorr
`Floben Alexander Hegole
`Jay Heineoke
`Mecnhard Herlvn
`
`Blostalisticn'in
`
`Peter Klein
`William M.F. Lee
`Edward E. MOITISBV
`Ellen Pure
`Daniel J. Flader
`
`Phyllis Gimottv
`
`Stevorr L. Fternor‘
`Amita sehgal
`John Stanleyl
`Doris Blotters
`Jellrey Weiser
`
`Katherine A. High
`Joy Hirsch
`Helen H. Hobbs
`Steven M. Holland
`Michael Holtzrnan
`Lawrence I-lolznian
`Jay Horton
`Goklran 8. Holamlsligrl
`Steven Houaor
`Mulseali Jain
`Carl H. Juno
`Takashr Kadowakr
`Fiaghu Knlluri
`Gerard Karsenly
`Flohert S. Kass
`Masalo Knsuga
`Daniel P Kelly
`Eli Kosher
`Surideep Khosla
`Flicharrl N. Kilais
`Thomas Fl. Kleyman
`Mark A. Kneppor
`Jav K Kolls
`loom Koniuro
`Murray Kort.
`Pant Kubcs
`Hohll Kulkarni
`l'laiiv liumal
`Thomas Krrrrper
`Anlnriin La Cava
`Mitchell A Lazar
`Rudolph L [Loin-I
`Klaus Luv
`Jan'ros t ion
`Polar LibbyI
`David Lillicrap
`Richard M. Locksiev
`Joseph A. Lorenzo
`Andrew Fl Marks
`Jack Marlin
`Koujl Malsushirna
`Michael A. Malthay
`Elizabelh McNaIIy
`Shlomo Melrned
`Michael E. Monclelsohn
`Thomas Michel
`
`Frank Miedernu
`Jotlory Mollvroritrn
`Anthony J. Muslin
`Martin G. Mvors.
`Erlt‘. G. Neilson
`Harrv T On
`Gary Owens
`Michael Parmurmk
`Cain Patlerson
`Warren S. Pear
`Alessandra Parnir.
`Shiv Pillar
`Kornelia Polvak
`Alice Prince
`Pare Purgserver
`Bali Pulendran
`Ellen Pure
`Marlene Rahinovitch
`Vincent Hacaniello
`Shahin Ralii
`Elaine Flames
`Gwendolyn Randolph
`Mariusz Flalalczak
`Barbara Retrermarin
`Murei'lach P. Reilly
`Giuseppe Helnuur
`Nicholas Realrlo
`Darrell Rollins.
`Marc E. Flotlionherg
`Paul B FlUll‘Itrrarl
`Anil K HllE-lgl
`..lunrcl1i Sar'loshirna
`J Evan Siidler
`Floliort I. Salirstein
`JUSG‘AiatIt Sahel
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`Mohamed Savoqlr
`Davrd Soho-inborn
`Cllrisliari Scliirirjlor
`Dollnl O. fichlondorll
`Michael D. Schneider
`Michael W “Schwartz
`John Sorlrir
`Randy Seal-fly
`Citrlh‘ilrtt-l Sorrlninn
`
`Flalick-Plerre Sakaiy
`Clay F. Semenkovich
`Gregg L Semenra
`William Sessa
`John Scykora
`Steven D Shapiro
`Kumnr Sharma
`Dean Sheppard
`Ethan M Shevaoll
`Mark Shirmrchlk
`Gerald Shulrflil"
`Roy L Silveré‘rlel“
`M. Coleste Slntoll
`LOIS Smith
`Weiliong Song
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`Warren Str'ohor
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`lanai;
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`Snow 8. Tliornorrsnr'rri
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`Carol lrrrv
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`Mallltlaa volt l-trrirnlll
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`

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`

`

`Contents
`
` Photo credit: Photo Researchers Inc.
`
`Cover: Colored scanning electron
`micrograph at an adipocytc. IWhite adipose
`tissue plays an important role in regulating
`metabolism. and its inflammation is
`linked to insulin resistance. Wueesl and
`colleagues report that expression ot the
`rteath receptor Fas is elevated in adipose
`tissue in genetic and nutritional models 01
`obesity in mice and in the adipose tissue ol
`patients with obesity and type 2 diabetes,
`and show that Fee is a regulator of obesity-
`associaten adipose tissue inllammalion.
`hepatic sloalosis. and insulin resistance
`{page 191}.
`
`Al
`flier
`
`Thejournal of Clinical Investigation
`
`Volume 120 Number 1
`
`January 2010
`
`'
`g
`.
`TR l O’M 3H
`l
`
`.
`H H
`r
`.
`I pf
`
`|
`l
`
`In this issue
`1
`
`r
`-
`l Editorial
`--
`H EAR—l— l
`2
`LEi'S keep this brief
`U.S. Nun
`
`
`
`tilt‘ 3510'” m EiTATIEEi
`
`: News
`
`Book reviews
`4
`Every patient tells a story: medical
`mysteries and the art of diagnosis
`D.W.Fosren
`Triumph Of "13 heart:
`Eh: story of statins
`.
`IBHY
`
`5
`
`.
`
`..
`
`..
`
`.
`
`-
`
`3
`
`Australian scientists say
`“G’day USA"
`K. Hourv
`
`
`
`stem ca"
`therapy for
`"eumdeseneraflve
`““39”"
`
`Review series Stem cells
`
`a
`
`11
`
`20
`
`29
`
`41
`
`51
`
`60
`
`71
`
`Stem cells: roadmap
`to the clinic
`[3.0. Deter
`
`Repairing skeletal muscle:
`regenerative potential of
`skeletal muscle stem cells
`F.S. Teossco. A. DELaniLLE,
`J. DIM-MANERA. G. Measure. into G. Cossu
`
`Pregenerative medicine:
`developmental paradigms In
`the biology of cardiovascular
`regeneration
`BA. Yr. 0. Wanner, mu KR. CHIEN
`Stem cells In human
`neurodegenerative
`disorders —- time for clinical
`translation?
`0. LiuoyALL AND 2. Koran
`
`The therapeutic promise of
`the cancer stem cell concept
`N.Y. anuir. T. Scunnon.
`
`“"0 M‘H‘ FRANK
`Progress toward the clinical
`application at patient-specific
`plurlpotent stem cells
`E. KISKINIS AND K. Eoom
`
`Enabling stem cell therapies
`through synthetic stem cell—
`niche engineering
`R. Pernnur AND RW. Zaunsrnii
`The bioethics 0131.9!“ cell
`research and therapy
`I. Hrun
`
`Thujuurnai oFClinical investigation
`
`itrrpr -'\\'\i'il'.1t'i.oi)1
`
`\‘niullu‘ 1?."
`
`Number I
`
`_i.ltili.!l'_\' _’l] ltl
`
`

`

`Cr.) ['1 it} r11" 1-5
`
`
`
`103
`Oncogenic m'I'OFI
`signaling
`
`
`
`1 27
`Cellular adaptation
`to hypoxia
`
`
`
`142
`
`Commentaries
`
`78
`
`80
`
`34
`
`87
`
`Putting the brakes on BTLA in T cell—mediated cancer immunotherapy
`CM. PAULUS AND CH. JNNr
`a Relalerlartlclc. page 15?
`Sodium channels gone wild: resurgent current trom neuronal and muscle
`channelopathies
`8.0. CANNON ANN E.P. BEAN
`Relaled article. page 369
`
`A‘
`
`New roles for Notch in tuberous sclerosis
`W3. PEAR
`Relaled arlIcles. pages; E33 and 103
`
`Oxidant stress derails the cardiac connexon connection
`G.F. TNNAar1LLI
`Related article. page 266
`
`N
`
`N
`
`90
`
`Flamping up HANTES in the acute response to arterial injury
`T. HLA ANN ML. ON
`a Related article. page 303
`
`Research articles
`
`93
`
`The evolutionarily conserved TSCIFlheb pathway activates Notch in tuberous
`sclerosis complex and Drosophila external sensory organ development
`M. KARBDWNICEEK. D. leSlltMNN. D. KNABINNN IN. T. HAINNIAN. J. Yu. T. MonntseN. E. NlceIAs. R. SNIINIAer.
`F. FlNEGIrns. ANN E.P. HENSKI
`a Related Commentary. page 84
`103
`Mammalian target of rapamycin regulates murine and human cell differentiation
`through STATsr‘pBSIJaggeleotch cascade
`J. MA. Y. MLNG. D.J. KwIArKNwsm. X. CINN. H. PFNG. [1. SNN. X. ZI-IA. F. WANG. Y. WANG. Y. JlNG. S. ZHANc.
`R. CHEN. L. WANN. E. Wu. G. CAI.
`l. MAI lNUWSKA-KULUIJEIH. D. LIAN. Y. LIN. Y. ZI-IAN. 0. SUN. K. Xu. J. DAI.
`J. HAN. L WN. HG. ZIIAN. H. SIIrN. AND H. ZNANN
`a Related Bummentary. page 84
`1 1 5
`Ablation of CIEBPB alleviates ER stress and pancreatic B cell tailure through
`the GRPTB chaperone in mice
`T MN':ll|N‘\. Y. KINN. S Afimlnlln. T. KAISlttl. T. TANAIIA. N. HABlllMtllfl. Y. SNINHAMA, A. TANtNA, T. INNNI.
`Y. SNINNIANI. M. KNYANAIN.
`l_ HIN;I'N1KA. M. MANrNMNIN. H. INNNE. T. UCI-NDA. M. Kern. Y. UCNIYAMA.
`S. ANINA. ANN M. KANNNA
`
`127
`
`The unfolded protein response protects human tumor cells during hypoxia
`through regulation of the autOphagy genes MAPtLC3B and ATGS
`KMA Htill'Lfliltli‘.
`I
`'Jlil‘l
`[II N Billlel N.
`l. Dl.tlitlIS.H. NtlS‘;lll,J. BUSSINK,K. SNUEI.KULJL5.T. KINerus,
`l-l_ MN ll.|t.l w Ll’IlillllYl. .JW. Villiljkl N. P.
`l AMINN. A..J. VAN [Jtli KNNlir . M. KflllllleSK‘I’, ANN BG. WNINI'IN:
`
`Targeting fatty acid
`oxidation in leukemia
`
`142
`
`Pharmacologic inhibition of fatty acid oxidation sensitizes human leukemia
`cells to apoptosis induction
`l. SAMIIIHIL ll. HI‘IIerIHril :, M. Tu l'.|
`. H. KAI-NAIIIIAN, M. KIJl-ltll‘l l‘m. B. Killlulll‘l. K. KAI NANANINNI.
`W. BNNl-IMANN. S DIIWNIN, ll. TAI MAN IN N_ ANN M. ANNNH ll
`
`
`
`254
`Cardiac fibroblasts
`and pressure overload
`
`15 7
`
`BTLA mediates inhibition of human tumor-specific CD81 T cells that
`can be partially reversed by vaccination
`L. Dr Hill. .J.-P. Hi"Ml‘-;, [j JNJlJll‘;. S. PNHUH. D. HIMNI NI.
`D. 0| Ivl'. ANN T}.F.. Set I‘ll N
`a RelaleIl Cmnmentary. page i5
`168
`CD27 sustains survival of CTLs in virus-infected nonlymphoid tissue in mice
`by inducing autocrine IL-2 production
`V. Pr-PrnrAK. Y. )(IAN. E.A.ll.ll. UrNAAN. ANN J. firmer
`
`I". FtNNN till, 0. Mililllll IN.
`
`179
`
`Lnk regulates integrin txlleS outside-in signaling in mouse platelets. leading
`to stabilization of thrombus development in vivo
`H. TAANAwA. S. NISNIMNHA. N. TAKAYAMA. A. DNA. H. NIFllllitll. Y. MNINIA. S. KAKN-INMA. S. YAMAMNI.
`S. OKAMNNA. N. TANNNA. 8. Gem. A. SAwAIrNerlr. l. MANANI . K. TNKNIFLIJ, ll NAAANNII. S.
`INKI’IKI. ANN K.
`
`I: it:
`
`'I‘IwJuurnnl quIiniml Investigation
`
`Imp-
`
`'uu'u- Ill III_I-_
`
`'v'uIIJmI- l_’.t:
`
`NIImI-rr
`
`|
`
`I.|||||.tl\ .'llItJ
`
`

`

`191
`
`203
`
`Deletion of Fas in adipocytes relieves adipose tissue inflammation and hepatic
`manifestations of obesity in mice
`5. WIJELSI. RA. RAPDI D. DM. SCIIIIMANN. JM. RYIKA. A. SCIIILDKNECNT. 0. Nov. AV. CHLRUUNSKY. A. Rumtzu.
`E.J. Scuneri. MY. DDNAIH. AND D. KDNDAD
`
`Urea-induced ROS generation causes insulin resistance in mice with chronic
`renal failure
`M. D'APDLIID. X. Du. H. ZDNn. A. CMIJCCI. L. MAIuAi. T. TRIUISANil. M. PriiDEuD-MANIDvANI. A. CAMPANDm.
`V. RAIA. J.E. Pr—ser. M. BHOWNLEE. AND I. GIARDINO
`
`214
`
`0020 deficiency in humans results in impaired T cell—independent
`antibody responses
`TW. KUIJPEItS. R.J. BFNDE. PA. BAAns. A. GNUMMELS. LAM. DEAAs. KM. DDLMAN. T. BEAUMDN‘I.
`T.F. TEDUEH. C.JM. vAN NO[SEI.. E. ELDLHING. AND RAW. vAN LIFR
`223 Genetic and epigenetic silencing of SCAHAS may contribute to human
`hepatocellular carcinoma by activating FAK signaling
`J. HUANG. D.'L. 2mm. F.-8. OIN. N. CHFNG. H. CHEN. B.—B. WAN. Y.—P. WANG.
`H.—S. XIAD. AND Z.-G. HAN
`
`242
`
`254
`
`266
`
`Cardiac mast cells cause atrial fibrillation through PDGF-A—rnediated fibrosis
`in pressure-overloaded mouse hearts
`C. LIAD. H. AAAzAwA. M. TAMAGAWA. K. Im. N. YASUDA. Y. Kunn. Fl. YAMAMDI'D. Y. UEASA. M. FUJIMDTO.
`P. WANG. H. NAKAUCHI. H. NAKAYA. AND I. KDMURD
`
`Cardiac fibroblasts are essential for the adaptive response of the murine heart
`to pressure overload
`N. TAKL'DA.
`I. MANADE. Y. UCHINU. K, EGUCHI. S. MAISUMDTD. S. NISHIMURA. T. SHINDD. NI. SAND. K. UTSU.
`P. SNIDEH. SJ. CONWAY. AND Fl. Mom
`
`Limited forward trafficking of connexin 43 reduces cell-cell coupling in stressed
`human and mouse myocardium
`JW. SMYTH. T.-T. HDND. D. GAD. JM. VDDAN. B.C. JENSEN. T.S. FUNG, P.C. SIMPSON. D.Y.R. SIAINIEFL
`NC. Cm. AND HM. SHAW
`
`7I
`
`Related Commentary. page 8?
`
`280
`
`290
`
`Cardiac signaling genes exhibit unexpected sequence diversity in sporadic
`cardiomyopathy. revealing HSPB7 polymorphisms associated with disease
`S.J. MAIKUVICH. D.J. VAN BDDIN-N. A. HINUES. NLY. KAND. T.E. DHULLY. ELM. VALLANIA. RD. METRA.
`MP. Fltu 1v. T.P. CAPPDLA. AND raw. 0an II
`
`TGF-Bt—induced expression of human Mdm2 correlates with late-stage
`metastatic breast cancer
`8. AAAKI. J.A. EIIEL. C.N. BATIJLLLD. K. BIJANGI-VESHEHSAHAEI. X.-J. XIE. D. DANIELPDUD. K.E. POLLUK.
`D.A. BOUTIIMAN. AND LD. MAYO
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`303
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`Stat3-dependent acute Flantes production in vascular smooth muscle cells
`modulates inflammation following arterial injury in mice
`J.C. KOVACIC. Fl. GDPIA. AC. LEE. M. MA. F. FANG. C.N. TDLDEm. AD. Wm 15. LE. BELmAN. H. SAN.
`G. CHEN. C. Si. HILAIRE. AND M. BUEHM
`N Related Commentary. page 90
`
`315
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`331
`
`343
`
`357
`
`The FlhoiRac exchange factor Vav2 controls nitric oxide—dependent responses
`in mouse vascular smooth muscle cells
`V. SAquAu. MA. SHNHA. M.J. MDNIEAD. AND KR. BUSTELD
`
`IL-‘l? produced by neutrophils regulates IFN-Y—mediated neutrophil migration
`in mouse kidney ischemia-reperl'usion injury
`L. LI. L. HuAND. AL. VERGIS. H. YE. A. BAJWA. V. NAHAYAN. RM. STRIETFR. D.L. RUBIN. AND MD. OKUSA
`
`The cytolytic molecules Fas ligand and TRAIL are required for murine thymic
`graft-versus-host disease
`I.—K. NA. 5.x. Lu. N.L. YLM. G.L. GOLDBERG. J. TSAI. U. RAD. 0M. SMITH. C.G. KING. D. SUN.
`0. HINSCHHDAN-CruEAMAN. L. PALDMDA. 0. PENADK. AM. HDLLAND. RR. JEND. A. GHOSH. H. TRAN.
`T. MFRGHUUB. C. LILI. G.D. SEMPUWSKI. M. VENTEVUGEL. N. BEAUCHEMIN. AND MRM. VAN DEN BRINK
`
`Foth expression in osteoblasts regulates glucose homeostasis through
`regulation oi osteocalcin in mice
`M.'T. RACHED. A. KDDE. B.C. SILVA. D.Y. JDND. S. GRAY, H. 0ND. J.—H. PAIA. RA. DLPINIIU. J.K. KIM.
`G. KAnsmrv. AND S. KUUSIFNI
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`commits
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`266
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`metastatic brew“
`cancer
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`315
`Regulating arterial
`contractility
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`Thcllourllai nI‘CIiIIical Investigation
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`369
`
`Human voltage-gated sodium channel mutations that cause inherited neuronal
`and muscle channelepathies increase resurgent sodium currents
`B.W. JnHEuKL. AD. Plumnz. J.0. JACKSON ll, nun TR. CIIMMINH
`Relaled Cmnrnentary. page 80
`
`S. Amnn, B. SUKI. AND DN. Kmltm
`
`Technical advance
`
`379
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`Amelioration of emphysema in mice through lentiviral transduction of long-
`lived pulmonary alveolar macrophages
`AA. WIlSDN. G.J. MURPHY. H. Hmnmwn, LW. KwoK. S. Snmwmsnm, A.-H. Hnww. RC. Muumnu,
`
`357
`Home control of
`mucosa home°5t35i5
`
`supplement
`390
`2009 American Society for Clinical investigation Presidential Address
`Can we keep the “academic" in academic medicine?
`N.C. Awumws
`
`Article amendments
`
`394
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`379
`Gene therapy for
`emphysema
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`iv
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`I'ti.'l.rti.-il Uriiiimiiriiniy. iilltill it: all
`
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`
`BTLA mediates inhibition of human
`
`tumor—specific CD8+ T cells that can
`be partially reversed by vaccination
`
`Laurent Derré.‘ Jean-Paul Rivals? Camilla Jandus,‘ Sonia Pastor,3 Donata Flimoldi,"
`Pedro Flomero.‘ Olivier Miohielin.‘-4 Daniel Dilute}5 and Daniel E. Speiser1
`
`'LllilWll} Institute in: Cancer Research. Lansanne. Switzerland. :‘UriiVGl'SliV Hospital Center fllltl llliwtifslly til tuiiszinne iGlIUVl.
`l.ElltS£ill|lll_ Swnrerlzinil. JINSICRM UMR 891. liistitiil Ptti‘ili Calmeltes, Marseille. France.
`"Miiltiilismnlinury Oncology Center. Laiisziiinii University Hospital (CHUVL Lausanne. Switzerland
`
`The function ofantigenwspecific CD8‘ T cells, which may protect against both infectious and malignant diseases.
`can be impaired by ligation oftheir inhibitory receptors, which include CTL~associatcd protein 4 (CTLA-4) and
`program med cell death 1 (PD-l ). Recently, B and T lymphocyte attenuator (BTLA) was identified as a novel inhib-
`itory receptor with structural and Functional similarities to CTLA-4 and PD- 1. BTLA triggering leads to decreased
`antimicrobial and autoimmune T cell responses in mice, but its Functions in humans are largely unknown. Here
`we have demonstrated that as human viral antigen—specific CD8" T cells differentiated from naive to effector
`cells, their surface expression of BTLA was gradually downregulated. in marked contrast, human melanoma
`tumor antigen—specific effector CD8‘ T cells persistently expressed high levels ofBTLA in vivo and remained
`susceptible to functional inhibition by its ligand herpes virus entry mediator (HVEM). Such persistence oFBTLA
`expression was also found in tumor antigen—specific CDB+ T cells from melanoma patients with spontaneous
`antinimor immune responses and after conventional peptide vaccination. Remarkably, addition oprG oligo-
`deoxynucleotides to the vaccine formulation led to progressive downregulation ofBTLA in vivo and consequent
`resistance to BTLA-HVEM—mediated inhibition. Thus, BTLA activation inhibits the function ofhuman CDS‘
`cancer-specific T cells, and appropriate immunotherapy may partially overcome this inhibition.
`
`Introduction
`
`.3 clitsscs ol signals:
`Atlivnlion til- li-“Iiipltoci'irs is :‘oltrl'olli‘tl l‘f.‘
`Iii si. hr 1 host- triggered through [hr '1‘ ccll rrccpt'or upon interac-
`tion with illlllIL'lt'l'Ilt' pcpl idu- l‘iilllltl to Ml Ii ' molecules; itiitl second.
`liti signal}. deliver-rd hr binding ol'corirci.‘ptors to their ligands on
`:tntnrvn prcscnt mg cells { l J.
`[ Tm'rct‘prot's consist of costiniiil.il'o
`r)! and ioiuhihnortr ii-ccpl'ors l2 7'}. Preclinical and cliniuil clam
`unlit-.m- ill." the l ii-ilil]il‘illil'}‘ receptors (ills-associated protein -l
`t'l' "I'l.r\--l] and pi ogi .illtl‘lll‘i‘l crll clcilth l (l‘l l- 1 l tire co—I'cspiinsil'ilc
`lot Ihc suppression of human clli'clor T cell responses i'o inlcc-
`[ll iii-i discurs'cs .‘llltl cancri l5. la}: the iln-rapeut‘tc liloclmde ill-llll'h-L'
`J p.il intuit“. r-. In promising clinical development. l.\-'mphocr-'tesrim
`csprc-is .nliliiiou.|| llllllllliill'l’ rcccpi'ors. such its k1llt‘l'-Il'lllll‘1ll‘l'l’
`m t-proi's .iud (.'-lcctin tt-pi- rccrptors till
`both of t hcsc classes.
`In ni-‘cvcr. .‘n'r csprcsscti Iii onlysmall stillsetsollcclls[8.81].
`A iiiol'c recently dcscrilicil L'tlvlllllll‘lllll')’ recrploi' Is ii and 'l' lyin-
`phocr‘ic attenuatorlll'l'l.:\; t ‘l JJTJ}. .in :itiiniinoglohulin-liltc mol-
`l't tilc belonging lo the t
`'l 128:“? family. which is expressed hi- the
`omit-citi- oi i}-'lill‘llll.!t._\'li’.‘i (is. it!
`ill. interestingly its ligand. hcrpcs
`\-'l| ll." cut 1'\' mediator [l l\"liM}. is a member of tile 'I‘NI‘ rccrplor
`l'l 'Nl’lil .-:tipct llnnilr l Hi.
`| I] This rccci'sior st'si'i-m also includes l_\'l‘tl-
`pltolosiu-(M It ill l'll"l)3.‘i.‘-§i..intl Ll}liil.l..\\'hicli.treprcsent .1! the
`t _\-'lIipl.isII1h inrmliranc ottirlls oldiil'crcru histological origin.’1‘hry
`m.“- ”unpro- lor tlicir ligand } WIN. \\'l'lli.'l‘l is picst‘nt on '1'. Hand
`.‘\ll\'
`L i‘ll.s. l N
`.'uic| tm-cloid cells. and also .i \'.ll'll_'l_\' ol‘l uniol‘ tells (it.
`Ill
`l3} l'lic ligation oi
`ll l'|.i\ l‘_\
`|i\'l-'l\l leads to pltospliorrlnlion
`ol i1iiinunort-ct-plor tt-rosnic lust-d Inhibition morils {l'l'lMsl and
`
`'l llllul of II m-n-sl cit-.I -.
`('ullllitl ill-interest: |".,.- .llI' ll- 'l -- ii.i\c lll'l l.|| I'll llr.iI n- I ‘ i
`(.il.||im1 lilrlliisarlitlmi I in lime IJII l." "
`in" .j_‘_i|lll:
`._1.n- lil ll _-‘-||
`'I-llltl"l|
`
`Lii'c homology 3 (Si l2} domain containingplior.pli.ll.‘isr l [HIP-1);"
`Hi I l’-3 association. resulting in dccrciist-d 'T cell prohl'craiion and
`criokinc prtuiucl'ion l
`l 3
`l-l}.1n I’scclls. ll'l‘l..-'\rcgultitcsllcrllircep-
`tor signaling li_\-' rcdocitig rhc phospliori-‘lation oi wk. 1% ccll linker
`protein (llLNKl. .llltl pliospholipnsv t “(.1 l_|‘I.l Tin-ll { | 5).
`It and T cell development is normal in ll-l-lJ‘WlL‘l-lllt‘lll' rnicc.
`Mature li'niphocrirs. howci'cr. .li'r l'unctionallr .tlicrctl .md show
`rnhanct‘d generation of Int-mori- T cells .ind memory responses
`llhl. ll'l‘l.i\ Lilli-it'll‘llt't\|.1Hl‘i.llllh.l Io t-nhantc protection l-l'tltl‘l
`ntiirint- malaria [ IT] .md to aggravate experimental .'lllrutii'iliitlrtr
`cliceplt.ilom_\_-ulii is (
`l .3) and .‘illrrgic anus!)- irill.iiiiiit.tii.iii t:
`| ti] .ind
`\\'.'t:s .‘issociiiivcl with spontaneous tit-i elopmcni ol'an .‘lllltlll‘nllllllll'
`hcpatitis liltcdiscnscl_l‘l). Mort-recently. ll'l‘l..-\li.1slirc|i shown to
`be involved in peripheral 'l'ct-ll i'oli‘r.‘incc induction {ill} and in only
`cont rol ol'tissuc damage and ill—Jill iluclcrinl immunity- {2 l ].
`in humans. li'l'l...-\ expression may be altered by specific immu-
`norhcrapr Will it allergens, as shown l'c-i' allergic rhinitis 1.12:. lln\\'-
`ever. only litrle is l(l'lU\|'l‘l about the mic and Iiinclion ol' ll'l'LA in
`humans. and there arc no data yet .tvailal’ilc on antigen-spec Mr 'I'
`cells. In this study. ire shmr tor the first time to our knowledge
`that ll'l'l_i\ is dou'nregtilaicd during l'tLli‘l‘ltll'l (TI 18' T cell tlillcrcn-
`nation to t-l‘lcctoi' cells. This was. lion-ever. not the case litl' tumor
`
`iiitrigcu- spccilh' erlan-AMW" ' specific} '1‘ cells. which pci‘sisirtii
`1}" cspi‘cs‘scd l’i'l'lA clcspilc cllccl'oi' ccll dillcrcntiat‘ion in uninc-
`L'llltllt’tl mclttnointi pririt‘nts. In contrast. win-n {ip( is were used .15
`adiuvani l'or vaccination, h-lt-lttii-l'V‘l‘l” '- specific '1' cells dou'ni'cgti-
`latcd ll'l‘l_.-\. tlt'Vl‘lI ipcti strongcllcctor functions. .ind lit-came indc-
`peiidcnt ot'l’s'l‘lA-mcdi.iti-d inhiliilion. l"iii.|ll_'_i'. the l-l'l'l.r’\ ligand
`l|\'l§i\'l tins lotmd to he expressed h}- melanoma tumors in sil u and
`ll'IL'Lllt'll't‘tl l'nticrionrrl il‘lllil‘ll'lt‘ll of ll'l‘l.r\'
`'l' cclls'.
`
`I'IIr- Iourlml til-Clinical Inresligaiinu
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`Imp
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`Figure 1
`BTLA [00272) is downregulated wrth progressive CD8' T cell ditlerenliatrnn, except in tumor antigen sponilrc (308' T cells Direct er: vivn
`analysis ol CD3' T cells lrom PBMCs by flow oytomelry. (A) Representative examples oi BTLA expression by Molan-AWH‘ ‘ and viriis-speoitlr:
`iEBV, CMV. and inlluenza virus [FLUlt (308' T cells from healthy donors (HD) and melanoma patients. BTLA' T cells- were distinguished lrom
`BTLA' T cells using a threshold established according to the aulologous naive T cell subset, which is always BTLA pnsrllvo. Numbers Indlcale
`percentage of positive or negative cells. {B} Comparison ol BTLA expression on total CD8' and virus- and Melan—A specilin loolls. Irom healthy
`individuals and unvaccinated melanoma patients. "P < 10
`for each of the 2 populations ol MelarI—A—speoitic T cells compared with all [a other
`populations. (C—E) BTLA expression in subsets of CDB' T cells of healthy individuals in -_ 58]. upon definition oi early and late dillerenliation
`stages by CD45RA and OCH? expression (Le. naive [N. CDdSRA'CCR'Z'L central memory [CM 0045RA CCH'Y']. alloctor memory IEM.
`CD45RA' CCFl7 ]. and elleotor memory FIA' [EMRA. CD4SRA'CCH7 ]ce|ls). gated as shown in C; a representative example demonstrates
`the narrow positive peak observed in phenotypio naive cells. [D] Statistical assessment at BTLA' CDB' T cell subsets. {E} GMT-l data. showing
`results compatible with those presented in D. Overall GIVIFI was determined by including the whole range lrnrn BT LA—negative to -pr1s|tivot:allsi.
`as shown in C. GMFI data were normalized to values determined in aulologous naive CD8' T cells.
`
`‘58
`
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`

`I'I..:st!il|'|2"ll
`
`;-itlir_'.|t_+
`
`Results
`
`l-"trm- but not tintmr-spctttit l‘n‘t’tlifl‘i .'l‘ lo" l‘rel'ia'rirttt'ttt'titgttltttefilial. 'I'tt
`Inuesligatt'theespressiolt olllil l.r\ lly tumor-specific (hlt'ifll‘l'AM'HH 1)
`and Virus—specific (LUV, f 'MV. and Inf ltienm virus} human ( '[).‘i'
`'1' cells. we performed es \’l\’lt multicolor flow cytontet ry analysis
`on t’BMt .‘s l'roni healthy donors and untreated melanoma patients
`(Figure IA}. In healthy individuals, |\-lelan-r\-“-"”'I specific ("lll'i'
`T cells were predominantly l’i'l‘lA‘ lliigtlt'e Ill). with significantly
`higher |i'l‘l.r\ expression than on Virttsvspecil-u' 'l‘ cells or on total
`(TIJh‘ T cells. This could he explained by their differentiation stag—
`es, since Melan-A-‘lw'
`' specific T cells are almost always naive
`in healthy donors (Supplemental Figure l‘. supplemental matc-
`i'ial ayailahle online with this article: doi: lll.l I73flfil4llll7lll )5] l,
`in contrast to virus-specific (fl )8'
`'1‘ cells (23.). tIottespondingly.
`naive T cells are H'I'IA positive In the mutine system (11,24,25l.
`'l‘ltel‘efot'e. we studied li'l'lfi. expression on total (TI )8‘ 'l‘ cells, in
`relation to their dii'ft't'entiatton stage as defined by expression of
`t_‘(_‘|t7 and (‘1 HSRA (2!!) in 58 healthy individuals {Figure Lt." and
`ll). Almost all phenorypic naive T cells (N) and the majority of
`central-memory T cells (VIN-ll expressed Il‘l'l.t\. In contrast. effec-
`tor meinory‘l‘ cells (liM) and effector memory RA' T cells [lift/IRA}
`were frequently ItTl.i\ negative, demonstrating decreased IETLA
`expression correlating with (708' T cell clil‘l-el'el'ttiatiori,
`in many instances, ll'l‘la’t receptor let-'els displayed a pattern of
`continuous surfitt‘t- expression without a clear transition from
`negative to positive T cells. However. phenotypically naii'e (TDS’
`T cells were consistently positive. with a distinct narrow peak. We
`therefore used tllléi population as a reference to set the threshold
`for l’i'l'lA' cells {l’igure HI). h'llll‘t‘t!\‘t‘l', we also evaluated the data
`in IL‘I'Il‘l'i ol‘ geometric mean fluorescence intensity {GMFI} val-
`ues normalized to the [iMlil oftht‘ autologous naive (‘1 )H T cell
`suliset. The results (l'igttre lli] closely resen‘thled those based on
`the assessrttent of percentage of li’l'lfi' cells {Figure ID]. 'I'hlthi.
`using autologons naive [3].}8‘ '1' cells as the internal HTLA refer-
`ence allott'trtl consistent slandal‘tlization and normalization of
`
`both percentage and UHF! data.
`Analyzing I’llMtls of melanoma patients revealed that IHLA
`expression was high among Melan-AM'“” lspecific T cells.
`surprisingly even among men-titty and effector cells {Figure 1H
`and Figure 3, A and B). In contrast. total CD8' T cells showed
`tt-dtttt‘t‘l HTLA expression. with progressive cliliferel‘ttiatiot't to
`t‘lli‘t'tttl tells [liigllt'e Ell).
`|-'oi- ciainpti-hensiye immune cliatactetizarion, it is also impor-
`tant to int-estigate'l'cells ftom diseased organsftissues. 'I'herel'ore.
`“a. analyzed lymphocytes from metastatic lymph nodes directly
`py- \‘l\-‘U and assessed Ill-LA expression l‘y b1crl.1|t—,:\M'\l“'
`I
`- N‘].‘|L‘t'il'_lt' T
`cellsas found iii 13 of [8 normal lymph nodes and in (i oft'i tumor—
`Iltfiltt‘ated lymph nodes. Once again. the h-lelal‘t-AMAN '—specil‘ic
`memory and I'll-L't‘lt‘tl' '1' cells were largely [t’l‘LA' . and the remain-
`.”an t '| W.‘ T cells. bore significantly lower levels ofli-TLA {Figure 2.
`t
`' and It). 'l'lius. the expression ofthe inhihitory receptor BTIA
`was high liir h-ll'l;‘ll'l-:\'.‘I'WI ' specific 'l' cells both in circulation and
`ill