.
Fostering Communication and Collaborati
The nihCatalyst A Publication for N I H Intramural Scientists
National Institutes of Health Office of the Director a Volume 13, Issue 4 a July-August 2005
Translational Research Success Story 30+ YEARS IN THE LPD: Cytokines Yield Treatment Advances Students/Colleagues In Inflammatory Bowel Disease Honor Franklin Neva by Celia Hooper by Karen Ross ncreasingly successful treat- On May 1 6, NIAID Is Laboratory ofPara- ment of a devastating pair of sitic Diseases (LPD) honored its newly re- chronic diseases, Crohn’s dis- tired long-time leaderFranklin Neva with I ease and ulcerative colitis (UC), a symposium at which Neva ’sformer and current colleagues traced LPD's (and stands as an elegant demonstra-
Neva ’s) history andpresented ongoing re- tion of the bench-to-bedsicle co- search. Tide symposium was organized operation that NIH boasts. by LPD Deputy Chief Tljomas Nutman, Presenting the work of scientists who co-moderated with LPD Acting Chief in NIAID’s Mucosal Immunity Sec- Alan Sher. tion of the Laboratory of Host De- Neva served as chieffrom 1969 until fense, Peter Mannon and his sec- few J- Mannon, M.D. 1995 and then as head ofLPD ’s Section tion chief, Warren Strober, recently Gastroenterology on Opportunistic Parasitic Diseases. He described gratifying clinical officially retired in December of 2004, progress in treating two forms of becoming scientist emeritus—and is still Celia Hooper a familiarface around the lab. inflammatory bowel disease (IBD). Warren Strober (left) and Peter Mannon The pair spoke at NIH’s “Demysti- the gastrointestinal (GI) tract, primarily The Early Days fying Medicine” course in early May.* patches of the small bowel and colon. Bedside breakthroughs sprang from Ted Nash was As Mannon describes it, inflammation the emerging of as a Neva’s first re- concept IBD of the GI tract can lead to reactions in Kafkaesque variant dis- search fellow. on autoimmune other tissues, including eye, skin, and He joined the ease in which a hyperactive immune bone. Ongoing symptoms may include system relentlessly LPD in 1970, a and inappropriately formation of fistulae and fibrotic obstruc- reacts not to oneself, but to next year after Neva the tions, abdominal pain, nutrient malab- became chief. closest thing—the bacteria that make up sorption, fever, bleeding, oral ulcers, part of the gut flora. Crossfire from the In 1969, Nash kidney stones, arthritis, uveitis, and a said, there was Karen Ross ensuing battle—and attempts to inter- variety of skin lesions. vene havoc with a patient’s gut vety little interest Franklin Neva: —wreaks Data from 1990 put the direct cost of Scientist emeritus and other tissues. in clinical parasi- Crohn’s at more than $1 billion, with By modulating key regulatory tology at NIH—and the lab was the high toll, Mannon says, reflecting to reactivity physically scattered, with sections in cytokines tone down the of Crohn’s tendency to strike young adults the system, Strober’s lab Georgia and Hawaii, as well as in immune has and to recur. The disease and the side Bethesda. Moreover, the scientists been able to eliminate IBD in mouse continued on page 4 tended to focus on the biology of models of the disease—paving the way individual parasites rather than on for translation to humans. Early-stage CONTENTS 6-7 Postbac Posters trials offer some promise of remissions parasite-human interactions, recalled 1 /Pioneer Awards Nash, who is now head of the Gas- in Crohn’s and UC patients. Translation: Cytokines Spark 8-12 trointestinal Parasites Section. IBD Therapy Interest Group There was, however, a growing The Clinical Picture of Crohn’s Directory need for a coherent clinical parasi- Crohn’s disease, occurring in 40 to 100 Honoring 13 tology program because soldiers of every 100,000 people, is a chronic Franklin Neva Cartoon Commentary disease unfolding from inflammation of were returning from the Vietnam War On Class Distinctions Editorial: suffering from parasitic diseases. * Webcasts the are available at Guest of course 14-15 Neva’s early training prepared him
Guest Editorial from a Former Deputy Director for Intramural Research
The Music of Science Plays On
The guest editorial this issue is from Lance Liotta, Principle 5: Strive to be taken for granted. who launched The NIH Catalyst in 1993, during The best administrative support is invisible. The his tenure as deputy directorfor intramural research Laboratory of Pathology has been fortunate to have in the time of Bernadine Healy's NIH directorship. tireless champions working behind the scenes for It is onlyfitting, then, that the departing remarks he its scientific staff—handling the myriad complex delivered at his going-away party at the Cloisters personnel activities that come with the territory of May 11 he adapted to fill this issue's DDIR column. the largest lab/branch at NIH and training hundreds of collaborators in the art of laser-capture micro- typical statement by the departing person at dissection and proteomics tecnology. a going-away party is that this job has been A the best years of their career. I cannot make Principle 6: Publications and patents are not that statement .... this has been the only job I the best way to extend science into the future. Ralph Isenburg have had since medical school. I joined the Labo- The best way is to train and inspire the future Lance Liotta, at his going-away party ratory of Pathology as a resident in 1976, and be- scientific leaders and teach them to make your cause I loved this place so loved the theories obsolete. lab’s May 11, 2005 much, and and technology Our sum- people so much, I stayed ever since. mer student program has literally changed the lives of hundreds of former students who are now phy- Over the last three decades, I’ve learned much sicians and scientists. from my role models—my co-workers, residents, students, and colleagues. Here are some of the important principles.
When I arrived here as a resi-
Principle 1: It is not enough dent, I viewed the intramural NIH to believe in a dream. There with the deepest reverence—and must be someone who be- I still feel the same way. This place lieves in the dreamer. In the be- has truly been a scientific utopia ginning of my career, my family driven by creative opportunity and Dr. [Alan] Rabson believed in and freedom, all directed toward me—so I began to believe in the goal of public health benefit. myself and had the courage to Here, each day, you can wake up
take risks. In 1997, 1 was privi- to pursue a new idea, tiy out the leged to meet Chip Petricoin, idea, and let the data drive the and together we proposed science. Here you have the great- our dream of proteomics to est opportunity to take risks and NCI. The institute appreciated test grand paradigms. Take advan-
our dream and helped us create a new clinical tage of that opportunity and privilege, as I have. proteomics program. George Gershwin was asked, “What comes first, Principle 2: The best way to see your the music or the lyrics?” “The money,” he answered, dream realized is to give it away. Transfer referring to the funding from his patrons. In this the dream to other team members and train- time of serious NIH fiscal restrictions, many have
ees; let them internalize it, improve on it, and expressed concern that the music of science will
call it their own. This is the secret to germinat- be silenced. I am an eternal optimist. Even if we, ing a large number of ideas in one’s short life- as scientists, have to do more with less, a bold cre- time. ative idea is still the best means to gain Principle 3: Taking a risk is the best way to funding and move the gain security in science. Instead of choosing the science to help pa- popular, safe topic that is in vogue, but will soon tients. Do everything to
be out of vogue, it is smarter to get into new terri- protect that freedom. tory, explore beyond the campsite, and discover your own promised land. After all, we really know —Lance Liotta Professor Life Sciences less than 1 percent about how biology works. of and Co-Director Centerfor Applied Principle 4: Hire people who are smarter Proteomics than you and then get out of their way—but and Molecular Medicine continue to lurk in the and shield them background George Mason University from that distract them. I have been problems might College ofArts and Sciences fortunate to work with world-famous pathologists. Virginia Fairfax ,
2 — . July — August 2005
Commentary from the NIH Assembly of Scientists
NIH Assembly of Scientists Recaps Conflict-of-Interest Work, Sets Up Shop for Continuing Activities and Activism
he Executive Committee of the As- cruitment and retention and employee visions in the conflict-of-interest regula- sembly of Scientists (AOS) was freedom. tions and their future implementation. As Telected in February of this year IS Fourth, to prepare the way for in- a result of that meeting, we have chosen and has been working hard to repre- stitutionalizing the Assembly of Scien- an Executive Committee member to serve sent the views of NIH scientists. We have tists to address other important issues on the Implementation Committee, and worked with representatives of intramu- related to morale and the quality of life we look forward to helping ensure that ral laboratory managers, extramural NIH of NIH scientists—including papeiwork, new regulations are implemented in a staff, NIH leadership, and members of resources, and respect— the Executive fair, balanced, and effective manner that professional societies. Committee has devised bylaws and an does not overwhelm NIH employees organizational structure. with paperwork. The First Five Months: Throughout this process the Executive Aims and Accomplishments Changes in the Air Committee has been working with the The Executive Committee has Along with many other factors, includ- deputy director for intramural research; achieved four objectives, particularly in ing extensive consultations with and AOS representatives are now regularly response to the conflict-of-interest in- work by Dr. Zerhouni, these activities attending the Scientific Directors meet- terim final regulations announced in have contributed to a change in atmo- ings to enhance communication. attitude in February 2005 (see The NIH Catalyst, sphere and —from one which Special Reference Issue,
may have to the regulations during the receptivity to our position made it much held in October 2005, and the newly official comment period and to com- more likely that a revision approximat- elected members will begin serving municate them as well to other inter- ing our proposal would occur in the in November. ested parties. policy rather than the legal arena. In For consideration, contact the nomi- Third, the Executive Committee addition, the withdrawal of the lawsuit nating committee, headed by Cynthia proposed an alternative set of regula- has not foreclosed any rights. Dunbar (
Call for Poster Abstracts for 2005 NIH Research Festival
he 2005 NIH Research Festival is set for October 18 through Other events during this 4-day annual showcase of intramural T October 21. research will include cross-cutting symposia and poster sessions; NIH and Bethesda FDA/CBER investigators may submit poster special exhibits on resources for intramural research; the Job abstracts online at
Honoring Franklin Neva Cytokines Advance IBD Treatment continued from page 1 continuedfrom page 1 ternship, Neva spent effects of standard treatments—corticos- a year and a half (De- teroids, 5-aminosalicylic acid (5-ASA), cember 1947 to July and immunomodulators such as azathio- 1949) in Egypt work- prine and methotrexate—often make ing at the Naval Medical Research Unit- (Case Western Reserve University, Cleve- work impossible and life miserable. Up 3 on typhoid fever and schistosomiasis. land)— spent just a few years in the LPD to 74 percent of patients eventually need Three years of research fellowships during their training. surgery to clear gut obstructions and at Harvard followed—the first working Others, such as Eric Ottesen (Emory other complications. Repeated removal on rickettsiae with John Snyder and the University, Atlanta), were part of LPD of segments of the small bowel in turn next two in the lab of John Enders, for decades but then moved elsewhere. may lead to additional nutrient absorp- where he learned techniques of grow- Still others, such as Amy Klion, David tion problems. ing viaises in tissue cultures. At this time, Sacks, and Tom Wellems, are current he described an exanthem disease (Bos- NIAID researchers. The Clinical Picture of UC ton rash) caused by an Echo virus. And representing two of the many sci- Similarly disabling, chronic, and re- After an academic appointment in the entists the world over with whom Neva lapsing, UC is an inflammation of the lab ofJonas Salk, he returned to Harvard collaborated were Barbara Herwaldt lower reaches of the bowel, from the to work in the newly created Depart- ( CDC) and Edgar Carvalho (Universidade distal rectum up into parts of the colon. ment of Tropical Public Health with Tom Federal da Bahia, Salvador, Bahia, Brazil). Continuous rather than patchy in its dis- Weller. It was here in the early 1960s The parasites they study include single- tribution, LIC affects only the lining of that Weller and Neva isolated and propa- celled organisms, such as Plasmodium bowel, without forming fistulae knot- gated rubella (German measles) virus. (the malaria agent), Leishmania, and ting into other tissues. Mannon says the It was also at Harvard that Neva ap- Cyclospora; and worms, such as schis- onset of LJC tends to be more abrupt plied tissue culture techniques to the tosomes, strongyloides, and filariae. than Crohn’s, but many of the general study of intracellular parasites and honed symptoms are similar, including diar- his clinical parasitology skills. Projects rhea, pain, and weight loss. Proctitic They work at the bench, at the bed- symptoms, such as rectal pain, urgency, The Neva Decades side, in the field, and everywhere in be- and bloody stools, are more prominent Upon his arrival at NIH, Neva con- tween. Weller studies the molecular func- in UC; affected tissues outside the GI solidated the LPD in Bethesda and em- tion of eosinophils, immune cells that system include the eye, skin, joints, and phasized research on the molecular bi- are critical for fighting parasitic infections liver. Patients are at risk for colon can- ology of parasites and the human im- but that also contribute to airway inflam- cer and inflammation of the biliary sys- mune response to parasitic infection. mation in people with asthma. tem. Two-thirds will develop chronic re- In 1971, he hired Louis Miller, currently Ottesen is working on a campaign to lapsing disease, and 40 percent will un- head of the Malaria Vaccine Develop- eliminate lymphatic filariasis by treating dergo surgery for UC. Current treatments ment Branch, who established the ma- the entire at-risk population in Africa include topical and oral 5-ASA, corti- laria research program. Before long, said with parasite-killing drugs. costeroids, and immunomodulators. Nash, “LPD was second to none” in clini- Herwaldt is a detective: She helped cal parasitology. pinpoint raspberries from Guatemala as From Mouse Models of IBD . . . Miller, whose malaria research was a the source of Cyclospora that sickened A view of IBD as an immune disor- key factor in LPD’s rise to preeminence, people in the United States in 1996. der began to emerge 30 years ago. In peppered his talk on malaria-host cell the past decade, Strober’s lab developed interactions with anecdotes about Neva Praise animal models that greatly refined that and the practice of clinical research in All of them talked about how Neva’s view, framing Crohn’s as a Thl-medi- the early days of LPD. insights and enthusiasm had inspired atecl immune disease heavily influenced their research. Weil, who works on the by IL-12, and LJC as a Th2-like disease People, Places, Parasites diagnosis and treatment of filaria infec- (like but not identical to a Th2-medi- The wide range, both geographic and tion, said he had Neva’s scientific spirit atecl response) heavily affected by IL- scientific, represented by the other sci- in mind when he developed an over- 13- In both, inflammatory cells in the entists who described their research at seas research program for Washington gut churn out their respective cytokines the symposium was a testament to the University medical students. in response to normal gut flora. impact that Neva has had on the field Sacks stressed Neva’s generosity, not- Gut bacteria were first implicated as of parasitic diseases. ing he was allowed to experiment with immune system provocateurs by the fact Some of them—including Peter Weller Neva’s precious collection of Leishma- that germ-free animals could not be in- (Harvard Medical School, Boston), Josh nia strains, even when he was a veiy duced to develop IBD, no matter what Berman (NCCAM), Gaiy Weil (Washing- inexperienced member of Neva’s lab. immune system genetic lesion research- ton University, St. Louis), David Free- Ottesen emphasized Neva’s leader- ers visited upon them. But IBD can be man (University of Alabama School of ship—of people and of projects. induced via gene manipulations in ani- Medicine, Birmingham), Chris Karp Miller urged Neva to continue to be a mals that harbor any of a large number (University of Cincinnati), Peter Melby frequent visitor. “The young people need of different gut bacteria or are exposed (University of Texas Health Science Cen- you,” he said, “and a few of us old guys to muramyl dipeptide, a component of ter, San Antonio), and Christopher King also love to talk to you." some bacterial coats. Strober says that 4 HHH July-August 2005
most of the thousands of antigens of the an antibody against 1L- gut flora are benign— but the species 12 in adult patients that can potentially trigger IBD “may with active Crohn’s dis- number in the hundreds and may vary ease. The multicenter, from individual to individual,” so that randomized, controlled, identifying the exact culprits for a par- double-blind trial of 80 Fig. 1-NOD2, a Negative Regulator of Peptidoglycan- ticular IBD patient is nearly impossible. patients included 63 Induced IL-12: NOD2 mutations lead to excessive IL-12- Strober’s lab used knockout mice to who received human induced Thl responses and the inflammation ofCrohn’s disease piece together the hypotheses that monoclonal antibody directed Crohn’s disease (see Fig. 1) involves vari- against the p40 subunit of IL- ous cytokines and NF-kB, NOD-2, and 12. The group tested two dose TLR-2, with dysregulation of IL-12, IFN- levels and two admin-istration y, and TNF-a. Strober's murine model schedules for seven injections of UC uses oxazolone (4-ethoxymethyl- of antibody. The treatment was ene-2-phenyl-2-oxazolin-5-one)—a con- well tolerated, and one of the tact-sensitizing agent applied to the rec- schedules for the higher dos- tum. Much like an adjuvant in a vac- age led to significant responses cine, the chemical spurs the immune and remission of disease com- system to respond to the flora that line pared with placebo (NEJM the lower reaches of the bowel. The 351:2069-2079, 2004). treated mice exhibit UC-like symptoms, Mannon was impressed that microscopic pathology, and changes in the patient response was as cytokines. good as or better than that with
Strober’s group was able to block coli- infliximab. He suspects it would tis symptoms by neutralizing IL-13 or be possible to obtain even bet- thwarting presentation of antigen ter results with refinements in through interference with CD1 on natu- dosing and maintenance regi- and Human Ulcerative Colitis ral killer T (NKT) cells. They now see mens. “It’s a very promising ap- UC as a Th 2-like inflammation with IL- proach,” Mannon says. The pharmaceu- ing of each patient’s dysfunctional im- 13 playing the lead role (see Fig. 2). tical company that owns rights to the mune relationship with his or her gut drug is examining application to other flora. Treatment would be targeted to
. . . To Clinical Trials of Cytokines diseases. NIH investigators have started identified patient subsets; a lab test
Crohn’s : Strober says studies of cells a pilot phase 2 study of G-CSF as an- would predict relapse risk, informing from Crohn’s patients confirmed that other approach to down-modulating IL- decisions for early intervention. immune cells in the gut mucosa secrete 12 and perhaps inducing T-regulatory increased amounts of IL-12 and other cells in Crohn's patients. Beyond IBD effector cytokines, including IFN-y and Ulcerative Colitis'. Investigational ap- Strober and Mannon believe concepts TNF-a. He believes IL-12, the master proaches to treating UC include: emerging from their work may have ap- cytokine of Thl T-cell differentiation, Dietary supplements and probiotics plication beyond IBD. Intervention drives the hyperactive immune response ingested to alter the gut flora against IL-12 — also at work in rheuma- in Crohn’s. Granulocyte pheresis toid arthritis, multiple sclerosis, and pso- Based on these and other studies, phy- Topical epithelial growth factor riasis —might be effective in suppress- sicians in the past few years have been Anti-IL-13 agents ing these other Thl immune-mediated using infliximab, an anti-TNF-a mono- Anti-CD3 and anti-IL-1 diseases. clonal antibody, to treat Crohn’s, enlarg- Strober says that one of the most Based on observations of Th2-like ing the percentage of patients with re- promising investigational drugs for UC, aspects of UC, investigators are looking missions and launching a new era of IFN-p, may work in accordance with his at the involvement of IL-13 in allergic increasingly focused immune therapies. lab’s model—by blocking NKT cell ac- reactions and asthma. But UC’s most Other agents under investigation prom- tivity, thereby curtailing IL-13 produc- important lessons may be more concep- ise to increase the frequency, durabil- tion. Mannon says the group has now tual, Mannon says, as it involves a dis- ity, and extent of response, while re- recruited more than half of the patients tinctive interplay of cytokines and cells ducing side effects, Mannon says. These needed for its pilot phase 2 study of IFN- at work. The tiny population of NKT include IL-10; ISIS-2302 (an antisense oli- P- A UC patient in the open-label trial cells that manage to cause such a large gonucleotide to ICAM-1); an anti-IL-6 describes prolonged periods without immune malfunction offers “a real op- receptor monoclonal antibody; thalido- disease relapse, and Mannon says the portunity to target therapy not broadly, mide; granulocyte and granulocyte-mac- early results are “very encouraging.” but to a very specific population of rophage colony-stimulating factor (G- Strober would like to see the devel- immunocytes that seem to be the bad CSF/GM-CSF); and anti-IFN-y. opment of a UC therapy based on more actors,” Mannon says, emphasizing the An NIH study group that includes direct, specific inhibition of IL-13. importance of focusing therapy on more Strober, Mannon, and Ivan Fuss com- Mannon sees the future of UC treatment upstream factors in the cascade of pleted a pilot phase 2 trial that tested in detailed, individualized understand- cytokines in immune responses. N 5 The NIH Catalyst
Postbac Poster Day. May 5, 2005: A Small Sampler by Aarthi Ashok HIV, Stroke, Dengue Among Targets of Postbag Research and Annie Nguyen
From Virus to Vaccine: Creating a She has been able to de- sible reasons for this sur- Live Attenuated Vaccinefor Dengue, sign two candidate vaccine prising finding. Type 2 strains: a dengue type 2 In future studies, the Caroline Agrawal, Boston University. strain with a 30-nucleotide candidate strains will be Preceptors: Steve Whitehead and deletion in the 3'-UTR se- injected directly into hu-
Joseph Blaney, MAID, Laboratory of quence ( D-30) and a 3- man liver tumor cells in Infectious Diseases nucleotide deletion in the SCID mice to assess their Dengue is a mosquito-borne virus NS3 viral protein that has level of attenuation. If with serotypes 1, 2, 3, and 4, which are both helicase and protease these candidate strains prevalent in most of the tropical areas activities; and a dengue show promise in the ro- of the world. Dengue causes a flu-like type 2/4 chimera that has dent model, they will be illness that can result in severe or fatal the structural protein of a used to vaccinate rhesus complications when a previously in- type 2 strain in a type 4 monkeys prior to chal- fected individual encounters a second- background in addition to lenge with wild-type vims. ary infection with a different serotype two point substitutions in The titer of neutralizing of the virus. No effective vaccines are the NS5 polymerase protein. antibody in these monkeys and the de- currently available. The ability of these candidate vaccine velopment of significant viremia or side Agrawal was interested in designing strains to grow in Vero cells, the intended effects will be closely monitored. If these a live attenuated vaccine strain of den- vaccine production cell line, was then strains are attenuated in monkeys, they gue virus in the laboratory primarily be- assessed. Although the type 2/4 chimera will be included in a tetravalent dengue cause of its immediate clinical applica- candidate grew efficiently in these cells, vaccine formulation to be used in hu- tions. She adopted a reverse genetics Agrawal and colleagues were surprised man clinical trials. approach, using PCR mutagenesis to to find that the type 2 D-30 strain with Agrawal plans to continue to study vi- modify the genome of a dengue sero- the single amino acid deletion in NS3 ruses during her graduate education at type 2 strain in order to attenuate the was unable to grow in these cells. Duke University in Chapel Hill, N.C. virus. Agrawal is currently exploring the pos- —Aarthi Ashok
RNase HI and Cel1 Viability: In vitro lar defects, Wirka decided HI in untransfected hu- RNAi Knockdown of Rnase HI to knock out RNase HI man osteosarcoma cells. Robert C. Wirka, University of Wis- function using siRNA. To Using RT-PCR and West- consin, Madison. Preceptor: Robert this end, he constaicted a ern blot analysis, he Crouch, NICHD, Laboratory of Mo- RNase Hl-GFP fusion con- hopes to determine the ef- lecular Genetics struct that was transfected ficiency of knockdown of Ribonucleases H (RNases H) are im- into human osteosarcoma RNase HI expression in portant enzymes required for the re- cells by electroporation. He these cells. moval of RNA in the RNA-DNA hybrids also co-transfected this con- RNase H is critical for formed during DNA replication in eu- struct with various anti- the production of infec- karyotic cells and during the conversion RNase HI siRNAs and tious HIV and is therefore of the viral RNA genome to DNA dur- quantified the knockdown an excellent target for ing HIV replication. In the latter case, of expression by FACS therapeutic drugs, Wirka the virally encoded reverse transcriptase analysis. notes. This research, he (RT) contains the RNase H activity. Wirka was able to obtain says, may inform the de- Wirka was interested in studying the specific knockdown of RNase HI ex- velopment of HIV RNase H drugs that function of RNase HI during normal cell pression using this system, with siRNA are extremely specific for the viral en- growth and proliferation. Previous ex- electroporation efficiencies estimated at zyme. periments in their lab had shown that 99 percent, RNase Hl-GFP knockdown “This has been an excellent experi- RNase HI gene knockout deprives mice at about 70 percent, and construct trans- ence for me,” says Wirka, who hopes to of the ability to replicate mitochondrial fection efficiencies at about 70 percent. remain involved in research during his DNA and is thus lethal. He plans to continue using and opti- clinical training at The Cleveland Clinic Because cells from these mice could mizing this siRNA strategy to knock Lerner College of Medicine. not be grown up to examine any cellu- down expression of endogenous RNase —Aarthi Ashok
NCCAM Launches Free Online CME Series CCAM has launched a new online Continuing Education Series in complementary and alternative N medicine (CAM). This user-friendly seminar series offers health-care professionals and the public the opportunity to learn more about CAM therapies and the state-of-the-science about them through video lectures by experts in their fields. Health-care professionals can earn CME credits; the course is free and can be found at
6 n July — August 2005
l
Glycodendrimers: Novel Previous studies have that glycodendrimers effectively inhibit Inhibitors of HIV-1 In- shown that glycoden- HIV-1 entry. fection drimers inhibit viral entry Although the mechanism of inhibition Benitra Johnson, South into host cells. remains unknown, Johnson offers two Carolina State Univer- Johnson carried out ex- possible hypotheses: There may be sity, Orangeburg. Pre- periments using a re- some form of modulation of the virus- ceptors: Robert Blumen- porter-gene-based assay cell interaction and/or there may be thal and Anu Puri, NCI, to measure the effective- some interaction between the glyco- Laboratory of Experi- ness of a novel glycoden- dendrimers and the host cell, leading to mental Computa- in altered cell physiology and abortion of and Annie Nguyen drimer blocking HIV-1 tional Biology Benitra Johnson infectivity. infection. Glycodendrimers are Successful viral entry Continuing glycodendrimer research, synthetic multivalent carbohydrate con- and integration into the host cell’s ge- she says, may lead to additional avenues jugates derived from analogous moieties nome leads to the expression of the lu- to counter HIV, which persists despite found on cell membrane surfaces; they ciferase reporter gene; quantification of the expanding marketplace of antiviral play a role in the binding interactions luciferase luminescence serves as an in- agents. of certain pathogens with mammalian direct measure of HIV infection. Prelimi- Johnson is contemplating applying to cells. nary data, Johnson reported, indicate medical school. —Annie Nguyen
Magnetic Resonance Imaging in Ex- The acute phase of stroke is charac- eas of cell death and cell swelling, de- perimental Cerebral Ischemia terized by cell death resulting from is- lineate actual lesions. Naomi Lewin, University of Mary- chemia, or decreased blood flow, either Lewin noted that the use of both MRI land, College Park. Preceptor: by direct or indirect causes such as techniques is better than either alone in Lawrence Latour, NINDS, Stroke apoptosis and inflammation. tracking and interpreting the course of Branch Two MRI techniques ischemia and cell death in the lab and The aim of Lewin’s re- were used to observe in the clinical setting. search is to develop neurological damage in Lewin’s lab aims to investigate poten- noninvasive MRI techniques rats. Perfusion-weighted tial markers of cerebral damage in rats to track cerebral changes in MRI using gadolinium and then translate the findings to hu- animal models of stroke contrast agent reveals man diagnosis. Ultrasmall superpara- throughout the course of light regions on the magnetic iron oxide particles are among stroke induction and recov- mean-transit-time map markers of inflammation under investi- ery. that signify decreased gation. Stroke is induced in the blood flow—which is in- Lewin plans to continue research in rat by inserting a silicon- terpreted to reflect areas neuroscience as part of an MD/PhD pro- coated suture into the at risk for damage. gram at the Weill Medical College of middle cerebral artery; the In addition, diffusion- Cornell University in New York City,
suture is removed after a Annie Nguyen weighted MRI, in which which she will enter this fall. given amount of time. Naomi Lewi light regions indicate ar- —Annie Nguyen
‘O Pioneers!’
he first annual NIH Director’s Pio- University, Waltham, Mass. (Mathemati- Rob Phillips, Ph.D., California In- T neer Award Symposium takes cal Modeling of Neural Systems) stitute of Technology, Pasadena (Ap- place on Thursday, September 29, George Q. Daley, M.D., Ph.D., Child- plied Physics) 2005, in Masur Auditorium. ren’s Hospital, Boston/Harvard Stem Cell Stephen R. Quake, D.Phil., The event will feature talks and a Institute, Boston (Stem Cell Biology) Stanford University, Stanford, Calif. roundtable discussion by the inaugu- Homme W. Hellinga, Ph.D., Duke (Bioengineering and Biophysics) ral group of Pioneer Award recipients, University Medical Center, Durham, N.C. Sunney Xie, Ph.D., Harvard Uni- selected in 2004, as well as the an- (Protein Design and Synthetic Biology) versity, Cambridge, Mass. (Cellular Im- nouncement of the 2005 awardees. Joseph (Mike) McCune, M.D., Ph.D., aging and Single Molecule Approach The day begins with opening remarks Gladstone Institute of Virology and Im- to Biology) by NIH Director Elias Zerhouni at 8:15 munology/University of California, San For the tentative agenda, see a.m. and ends with an informal re- Francisco (HIV Pathogenesis)
7 The NIH Catalyst
Interinstitute Interest Group Directory
Web Access
Although not all the sites are up notices by e-mail and on the SBIG website Bioethics Interest Group to date, nearly all the Interest Contact 1: Susan Buchanan Meeting time: 1st Monday (except 2nd Groups have web sites that can be Phone: 301-594-9222 Monday following holidays; usually does accessed through
Calcium Interest Group Cognitive Neuroscience Consortium Pittsburgh, PA; Univ. of North Carolina, Meeting time and place: Not regularly Meeting time: Every two months, last Chapel Hill, NC; Oregon Health and Science scheduled at this time Wednesday, 4:15 pm Univ, Portland, OR Contact 1: Arthur Sherman Meeting place: NSC Building, Conference Contact 1: Kenneth Kraemer Phone: 496-4325 Room A (starts September 2005; Extramu- Phone: 301-496-9033 E-mail:
Contact 1 John Schwab Contact 1 : Sabra Woolley Meeting place: Building 6B, 4B429 : Room Phone: 301-594-3827 Phone: 301-435-4589 Contact: Jim Kennison E-mail:
Phone: 301-594-8483 E-mail :
Meeting time: Monthly, last Wednesday, noon E-mail :
Interinstitute Interest Group Directory History of Biomedical Research Interest Group Meeting time: Second Tuesday, 1:00 pm ** Endocrinology Interest Group Genomics and Bioinformatics Interest Meeting place: Varies; check web site Meeting time and place: Varies Group Contact 1 : Office of NIH History Contact 1: George Chrousos Meeting time: Usually one Thursday' Phone: 301-496-6610 Phone: 301-496-5800 month, 3:00 pm Contact 2: Victoria Harden E-mail:
Contact 1: Ingrid Li Epidemiology and Clinical Trials Glycobiology Interest Group Phone: 301-443-1421 Interest Group Meeting time and place: Varies E-mail:
5N264 Contact 1 : Wendy Weinberg Meeting Place: NIBIB, 6707 Democracy Contact: Jay Knutson Phone: 301-827-0709 Blvd, Bethesda, Suite 200, Room 223 Phone: 301-496-2557 E-mail: E-mail : Lambda Lunch (Bacterial and Phage Molecular and Functional Pigment CeE Research Interest Group Genetics) Optical Imaging Interest Group Meeting time: Monthly, usually 3rd Meeting time: Each Thursday, 11:00 am Meeting time: 2nd Wednesday, 12:00 noon Thursday, 12:30-2:00 pm; yearly day-long Meeting place: Building 37, Room 6107/ Meeting place: Building 10, Room B3MB- meeting most years; check the website 6041 38 (2.5 level, B-wing) Meeting place: Bldg 49, Conf. Room 1A51 Contact: Susan Gottesman Contact: Amir Gandjbakhche Contact T. Marjan Huizing Phone: 301-496-3524 Phone: 301-435-9235 Phone: 301-402-2797 E-mail: Interinstitute Interest Group Directory Stem Cell Interest Group Tobacco and Nicotine Research Interest Contact 2: Alan Hinnebusch Meeting time and place: Monthly seminars to Group Phone: 301-496-4480 rotate through Baltimore, Bethesda, and Meeting time: 4th Wednesday, every other E-mail: E-mail: Washington Hospital Center Contact 2: Uli Siebenlist newsletter: Interest Group E-mail : E-mail: E-mail : 12 July — August 2005 Commentary Why a Postdoc’s Life Reads like a Bad Jane Austen Novel by Fatima Husain, nidcd (if she had written one) N Class distinctions rule: 1 hour of PI work = 20 hours of Postdoc = 40 hours ofPostbac; calculate authorship accordingly J r At conferences, the Pis congregate with other Pis and the Postdocs stand by their posters like so many wallflowers waiting to be asked to dance V. J V 13 The NIH Catalyst People Recently Tenured Kyung Lee received his Ph D. in 1994 of mammalian Wee 1 ) inhibits mitotic en- mosome missegregation and genomic in- from the Department ofBiochemistry at try by negatively regulating Cdc28. We stability, thus promoting tumorigenesis. the Johns Hopkins University in Baltimore. found that Cdc5 directly phosphorylates We expect that understanding Plkl-depen- He then worked with Raymond Erikson at and downregulates Swel,thus permitting dent PBIP1 regulation may provide new Harvard University in Mass., activation of and thereby mi- insights into Plkl deregulation Cambridge, the Cdc28 how pro- as apostdoctoralfellow and studied in the totic entry. The conserved polo-box do- motes cancers in humans and how to fields ofcellularproliferation and mitotic main of Cdc5 appeared to play a crucial approach the development of anti-Plkl controls. In 1998, hejoined NIH as a ten- role for this biochemical step by target- therapeutic agents. ure-track investigator in the Laboratory ing the catalytic activity of Cdc5 to its sub- ofMetabolism at NCI and is now a senior strate Swel. Francesco Marincola received his MD investigator and head of the Similar regulation occurs in from the University of Milan (Italy) in j Chemistry Section. human cells, where down- 1978 and completed a general surgery During the eukaryotic cell regulation of Wee 1 by Plkl is residency at Stanford University (Stanford, cycle, reversible protein critical for entry into mitosis. Calif.) in 1990. He joined the Surgery phosphorylation by protein These findings demonstrate Branch, NCI, in 1990 as an immuno- kinases is a fundamental the importance of the polo- therapyfellow and subsequently became regulatory mechanism. The box domain for specifically tar- a surgical oncology fellow. In 1993, he G/M phases of the cell cycle geting the Plks to their sub- was appointed to the staff of the Surgery comprise a series of regula- strates through protein-protein Branch; in 2001, he assumed two addi- tory biochemical steps and interactions. tionalpositions—director ofthe Immuno- coordinated cellular events Dynamic subcellular local- genetics Program in the Clinical Center that ensure the faithful parti- ization and the multitude of Department ofTransfusion Medicine and tioning of genetic and cyto- Plks functions predict that the director of the HLA laboratory. plasmic components. polo-box interacts with mul- Since my immunotherapy fellowship in It is now clear that, in addition to the tiple cellular proteins at specific stages of 1990 with Steve Rosenberg in the NCI much-studied cyclin-dependent protein the cell cycle. To gain a deeper under- Surgery Branch, I have been intrigued by kinases, the polo subfamily of serine/ standing of the function of the polo-box, the rare yet overwhelming observation of threonine protein kinases (collectively, we have been focusing on the dramatic tumor regressions polo-like kinases, or Plks) plays a pivotal identification of novel polo- mediated by immune ma- role in regulating various mitotic events. box-binding proteins in cul- nipulation with the systemic Plks cooperate with a master cell-cycle tured mammalian cells. administration of interleukin- kinase, Cdc2, to bring about essential mi- From a yeast two-hybrid 2 (IL-2) — with or without tu- totic events at mitotic entry; Plks also regu- screen using the polo-box do- mor-infiltrating lymphocytes. late pathways leading to the inactivation main of Plkl as bait, we iso- When the fellowship ended, of Cdc2 at mitotic exit. lated a novel kinetochore pro- I agreed to continue in the Loss or depletion of Plks activity in vari- tein that we termed PBIP1 staff of the Surgery Branch to ous organisms results in mitotic arrest fol- (polo-box interacting protein work on the identification of lowed by cell death, suggesting that the 1). Consistent with the two-hy- the algorithm regulating tu- function of Plks is essential for mitotic pro- brid analyses, PBIP1 interacted Fran Pollner mor rejection in humans ex- gression in all eukaryotic organisms. with endogenous Plkl in vivo Francesco Marincola posed to this kind of therapy. LJsing both budding yeast and cultured under physiological condi- As the tumor antigens (TA) mammalian cells, my laboratory has in- tions. recognized by lymphocytes became mo- vestigated the mitotic functions of Plks. Preliminary characterization of PBIP1 lecularly characterized in the ’90s,TA-spe- We have shown that the polo-box motif, suggested that it is a kinetochore-specific cific vaccines were developed that yielded which resides in the C-terminal noncata- mitotic inhibitor that is phosphorylated the unprecedented opportunity to study lytic domain, is essential for the localiza- and degraded by Plk 1 . Expression of the tumor-host interactions specific to a tion of both budding yeast polo kinase nondegradable PBIP1 lacking the Plkl- unique TA and in some cases to a spe- Cdc5 and mammalian Plkl to specific sub- dependent phosphorylation sites induced cific TA-human leukocyte antigen (HLA)- cellular structures. a mitotic arrest, indicating that degrada- specific combination. This ability to mini- In line with this notion, mutations of tion of PBIP1 is required for proper mi- mize experimental variables in humans the polo-box of either Cdc5 or Plkl in its totic progression. simplified the study of the effect of im- native organism abolished the function of Intriguingly, PBIP1 interacted with cel- munization in clinical trials. these enzymes and induced mitotic ar- lular proteins whose deregulation is im- We discovered that TA-specific immu- rest, indicating that polo-box-dependent plicated for tumorigenesis, suggesting that nization can reproducibly induce cytotoxic subcellular localization is critical for the the Plkl -dependent downregulation of T cells (CTL) in patients that are capable mitotic functions of these enzymes. PBIP1 may contribute to the development of recognizing tumor cells. In addition, One of the critical events that budding of cancers in humans. In line with these we observed that the CTL induced by vac- yeast polo kinase Cdc5 mediates is mi- observations, Plkl was both upregulated cines localized at tumor sites, where they totic entry, a process that requires activa- and multiply mutated in about 80 percent recognized tumor cells and produced tion of Cdc28 (homolog of mammalian of human cancers. cytokines such as IFN-y. Cdc2).We set out to clarify the molecular Our recent results raise the possibility Such interactions, however, were not mechanisms of Cdc5 regulation of this that PBIP1 is a novel tumor suppressor. sufficient to induce tumor regression. In event. Precocious degradation of PBIP1 by de- addition, we observed that the level of Studies have shown that Swe 1 (ortholog regulated Plkl activity may lead to chro- expression of TA or HLA molecules re- 14 — July-August 2005 sponsible for the presentation of the TA the direct ex vivo analysis of sively parallel signature se- to CTL did not predict response, suggest- tumor-host interactions during quencing, microarray, and ing that other factors modulate immune vaccine therapy. Our aim is to EST scan — developed at NIH responsiveness at the tumor site. identify factors predictive of and elsewhere to identify The development of high-resolution the immune responsiveness of novel genes and new signal- molecular testing of HLA molecules, us- patients and/or their tumors. ing pathways that regulate ing high-throughput sequencing technol- These tools include the com- self-renewal. Using such ogy, was critical in these early vaccine bined analysis of DNA, RNA, methods, we were able to studies and was made possible by Harvey and protein to identify genetic and epi- cross-correlate information to obtain a glo- Klein in the CC Department of Transfu- genetic differences between patients who bal view of cell function and start to iden- sion Medicine (DTM), who offered me a do and do not respond to therapy. tify context-dependent and cell type -spe- joint appointment as director of the HLA Using affordable, custom-made oligo- cific signaling pathways. For example, we Laboratory in addition to my original ap- nucleotide-based chips, we are develop- identified Lin4l as a microRNA-regulated pointment in the NCI Surgery Branch. ing high-throughput screening tools to gene that is important in regulating em- We employed a novel strategy to ana- analyze cytokine polymorphisms and test bryonic stem-cell self-renewal. lyze T cell localization at tumor sites dur- whether genotype can be linked to phe- In the past couple of years, we have ing immunization and the interactions of notypic responses and to clinical param- extended this strategy to examine human T cells with their target (the HLA-TA com- eters. We are also continuing to improve embryonic stem-cell populations— with bination): With multiple fine-needle-aspi- our proteomics and functional genomics equally exciting results. Most of our find- ration biopsies, we could monitor tumor- tools for the global assessment of patients’ ings have followed as a logical conse- host interactions in the target tissue ex response to treatment during clinical tri- quence of the key observation that all vivo while leaving the tumor in situ, al- als. stem-cell populations generate differenti- lowing us to follow the natural history of ated progeny by first generating more re- the disease and serially evaluate response Mahendra Rao completed his medical stricted precursors. to therapy. training at Bombay University in India in For example, based on our initial find- In 2000, Ena Wang in my laboratory de- 1983 and received his Ph D.from the Cali- ing, we were able to hypothesize and then veloped and validated a technique for fornia Institute ofTechnology in Pasadena confirm that repair in the adult nervous RNA amplification that allowed the use in 1991- Afterpostdoctoral training with system likely involves a larger role for re- of very small amounts of starting RNA (ob- Story Landis, then at Case Western Reserve stricted progenitors than for true multi- tained from fine-needle aspirates) in the University in Cleveland, and DavidAnder- potent stem cells. study of tumor-host interactions in con- son at the California Institute ofTechnol- This result has important implications junction with high-throughput technolo- ogy, he joined the Department ofNeuro- for both embryonic stem-cell and neural gies such as microarrays. This research biolog)' andAnatomy at the University of stem-cell biologists. It suggests both that expanded our ability to monitor immune Utah School ofMedicine in Salt Lake City stem-cell populations need to be differ- responses during immunization and other in 1994. In 2001, he joined NIA as chief entiated prior to implantation and that for immune therapies. of the Stem Cell Biology Unit in the Labo- specific diseases, subsets of cell types may These original studies suggested that the ratory of Neurosciences. He is currently be more important — glial for glial disor- tumors most likely to respond to immu- also an adjunct associateprofessor at the ders and neuronal cells for diseases that notherapy are characterized by an immu- National Centerfor the Biological Sciences include neuronal loss. nologically active (chronically inflamed) in Bangalore, India, and holds ajoint ap- We then proceeded with transplant ex- transcriptional profile. When successful, pointment as an associate professor at periments using rodent models of immune therapy — whether TA-specific Johns Hopkins Un iversity, Baltimore. Parkinson’s disease and spinal cord in- immunization or the systemic administra- At both the University of Utah and NIA, jury to show that these restricted precur- tion of immune stimulants such as IL-2 my laboratory has focused on understand- sors have a therapeutic benefit. This acts by turning a chronic inflammatory ing how embryonic and adult stem cells opened up an entirely new field of en- process into an acute one quite similar to self-renew and differentiate. Our early deavor, new collaborations, and interac- that observed by others in the context of work with fetal and adult neural stem cells tions with new colleagues. acute transplant rejection. revealed that they change over time with I have benefited enormously from the Subsequently, Monica Panelli, also in respect to their self-renewal capabilities, efforts and intellectual input of people in my group, compared fine-needle aspirates expression of markers, positional infor- my own laboratory and from Mark obtained before and during IL-2 therapy mation, and differentiation ability. Mattson and other colleagues in the Labo- and showed that this cytokine acts by in- We then showed that stem cells do not ratory of Neurosciences —and from NIA ducing a general inflammatory process at differentiate into fully mature cells directly and NIH resources and infrastructure that the tumor site. This inflammatory process but do so by first generating intermediate allow relatively small laboratories to un- activates innate effector mechanisms in precursors, which we termed lineage-re- dertake large-scale analyses. mononuclear phagocytes and natural stricted precursors. We subsequently iso- In the coming years, I hope to build on killer cells and induces activation of anti- lated specific lineage-restricted precursors the wealth of information we have gen- gen-presenting cells and production of and showed that they play an integral part erated to begin to understand how stem chemoattractants capable of recruiting and in the normal process of development. cells maintain their self-renewal capabil- activating T cells at the tumor site. Indeed, their action at specific stages can ity and avoid senescence. Further, com- In 2001, Klein invited my group to de- explain the effects of many genes. parisons across species will allow us to velop the Immunogenetics Program in the Our ability to identify specific popula- determine what common and/or distinct DTM. We are now developing high- tions of cells allowed us to use the large- mechanisms of self-renewal have evolved throughput, hypothesis-searching tools for scale analytical procedures— such as mas- over time. H 15 The NIH Catalyst Catalyst; Worth Its Weight m G®{4- Catalytic Eureka! Part 1 Reactions? ne day about 2000 years ago, the Greek King Heiro wanted a new crown. So he weighed f you have a photo or out some gold, gave it to his local crown-mak- I other graphic that O ing craftsperson, and soon had a shiny new diadem. reflects an aspect of life at But something made King Heiro suspicious. Convinced NIH (including laboratory he had been cheated out of some of his gold, he asked life) or a quotation that around for help on how to prove it. scientists might appreciate The problem eventually came to a very clever guy that would be fit to print in named Archimedes (you’ll hear his the space to the right, why name many times again in travels!). not send it to us via e- your Bright as he was, this one had mail: [email protected]>; him stumped. How to prove the crown wasn’t 100 percent gold? After days of contemplation, fax:402-4303; or mail: he finally took a break. What does a frustrated scientist do to relax? Take a bath. Building 2, Room 2E26. Archimedes stepped into his tub and the veiy same thing happened then as would happen now if you got into a tub full of water: It would overflow. So with water spilling all over the Also, we welcome place, the solution to the problem hit him. He was so excited that he was crying “Eureka!” “letters to the editor” for (Greek for “I have found it”) out the door, through the streets, and to the king. (In his glee, publication and your he apparently forgot his robe, guaranteeing that this would be the stuff of legend!) reactions to anything on So how did Archimedes figure it out? Here’s what he knew: the weight of the crown and the Catalyst pages. what the crown was allegedly made from. He placed the crown into a vessel full of water to see how much water overflowed. Then he did the same with the same amount weight-wise of gold. Even though the crown was the same weight as the gold, it displaced a different In Future Issues... amount of water. The king had his proof, the crown maker got his just reward, and Archimedes put his robe on. you don’t need gold crowns lying to test a Yoga Therapy Now around the volume of irregularly shaped objects, and you certainly don’t need anything more than a carefully-made scale to tell that two things shaped the same are made out of different materials. B CRADA Country Next issue we will take a bunch of pennies and prove that not all pennies are created equal using a surprisingly accurate home scale. Maybe you can work on that problem in the meantime, but here's a hint: Compare U.S. pennies made before and after 1982. You’ll be More Bench surprised! To Bedside —Jennifer White The NIH Catalyst is pub- Publisher Managing Editor Editorial Advisory Board lished bi-monthly for and by Michael Gottesman Fran Pollner Jorge Carrasquillo, CC the intramural scientists at Deputy Director David Davies, NIDDK NIH. Address correspon- for Intramural Research, OD Copy Editor Dale Graham, CIT dence to Building 2, Room Shau na Roberts Hynda Kleinman, NIDCR 2E26, NIH, Bethesda, MD Editor Elise Kohn, NCI 20892. Ph: (301) 402-1449; John I. Gallin Contributing Writers Susan Leitman, CC fax: (301) 402-4303; Director, NIH Clinical Center Aarthi Ashok Bernard Moss, NIAID e-mail: U.S. Department of Health FIRST-CLASS MAIL and Human Services POSTAGE & FEES PAID National Institutes of Health DHHS/NIH Building 2, Room 2E26 Permit No. G-802 MSC 0235 Bethesda Maryland 20892 Official Business Penalty for Private Use $300 ® Printed on 50% recycled content paper and can be recycled as office white paper.