first indo-us bilateral meet on molecular imaging

Indo-US Science and Technology ForumIUSSTF

Indo-US Workshop on

Applications of Molecular Imaging in Health, Disease and Drug Discovery

Nov 8-10, 2010

Centre for Cellular & Molecular BiologyHyderabad - 500 007, India

CCMB

arestream

Co-organized by

Indo-US Workshop on “Applications of Molecular Imaging in Health, Disease and Drug Discovery” 1

Imaging of molecules, cells, tissues, organs and whole organisms has been en vogue ever since the first X-ray images of Albert von Kolliker's ring bearing hand were taken by Wilhelm Conrad Roentgen in 1896. Over the past century significant advancement has occurred in this field, that has increased the sensitivity and specificity of biomedical imaging and completely changed the way radiological medicine is practiced. The first decade of the new millennium has brought even more impetus to the field thus beginning a new era for the application of “imageology” in a large spectrum of disciplines that includes interactions between small molecules, understanding higher order brain functions and discovery of new drugs.

The labs in United States have been pioneers in this newly developing science and the National Institutes of Health has started a new institute specifically targeted for research on biomedical imaging, whose Deputy Director is attending the workshop. Indian labs are also becoming aware of the power of these developments and new infrastructure is being established at several institutes where bioimaging will be practiced extensively. The Indo-US Science and Technology Forum, New Delhi recognized this synergy between the US and Indian labs and approved a proposal submitted by CCMB, Hyderabad, India and Carestream Health Inc. Woodbridge, USA to hold this workshop. On behalf of our respective institutions we are privileged to host a very knowledgeable faculty for the workshop and we warmly welcome all the participants to CCMB Hyderabad. We are thankful to the Director of CCMB who has graciously extended the use of all the facilities at CCMB for this workshop.

Hope all of you will have fun and learn new things in the two and a half days that you spend here!

Gopal Pande Rao PapineniCCMB Carestream Health Inc.

A Note from the Organizersto

ALL PARTICIPANTS AND SPEAKERS OF THE INDO-US WORKSHOP ON “APPLICATIONS OF MOLECUALR IMAGING IN HEALTH,

DISEASE AND DRUG DISCOVERY” NOVEMEBER, 8-10, 2010

atCCMB, HYDERABAD, INDIA


Venue: CCMB Lecture Hall

AGENDA

Indo-US Workshop onApplications of Molecular Imaging in Health, Disease and Drug Discovery

November 8-10, 2010



6.00 pm - Arrival and Mixer – CCMB Centre Court

thMonday, the 8 November, 2010

8.45 am - Opening Remarks by

Ch. Mohan Rao, Director, CCMB, Belinda Seto, Dy. Director, NIBIB and Nishritha Bopana, Science Officer, IUSSTF

Session I : Chairperson – Kakarla Subbarao9:15 to 11:00 am

Molecular Medicine: Present and Future Belinda SetoNIBBB, Bethesda

Potential of MRI and In-vivo MR Spectroscopy in Disease Process N.R. JagannathanAIIMS, New Delhi

11.00 - 11.15 am - Tea/Coffee Break

Session II : Chairperson – Arun K. Gupta11.15 - 1.00 pm

Pre-Clinical Molecular Imaging Applications in Cancer Research Rheal TownerAMRC, Oklahoma City

Use of optical spectroscopic and imaging techniques for biomedical diagnosis P.K. GuptaCAT, Indore

1.00 - 2.00 pm - Lunch Break

Session III : Chairperson – Kit Lam02.00 - 03.40 pm

Image guided interventions A. K. Gupta SCTIMS, Thiruvananthapuram

Combined Anatomical and Molecular Imaging with Therapeutics using Shanthi K. Nair Biocompatible Nanoparticles AIMSRC, Kochi

03.40 - 4.00 pm - Tea Break4.00 pm - Departure to Golkonda Fort

8.00 pm - Dinner at Taramati

thSunday, the 7 November, 2010



thTuesday, the 9 November, 2010

Plenary Session IV : Chairperson – Belinda Seto9.00 – 11.00 am

From combinatorial chemistry to nanoparticles to Kit Lamcancer imaging and therapy UCDCC, Davis

Imaging RNA in cells and in animals Samit AdhyaIICB, Kolkata

11.00 - 11.15 am - Tea/Coffee Break

Session V : Chairperson – Arvind K. Chaturvedi11.15 - 1.00 pm

Nano "Solutions" for Therenostics Mark KesterPenn State Univ., Hershey

Fluorescence based imaging of tumor cells Gopal KunduNCCS, Pune

1.00 - 2.00 pm - Lunch Break

Session VI : Chairperson – B. Ravindran02.00 - 03.40 pm

In vivo cellular imaging and its role in cell therapy Ali ArbabCMIL, HFH, Detroit

CNMR Investigations of Excitatory and Inhibitory Anant PatelNeurotransmission in Cerebral Disorders CCMB, Hyderabad

03.40 - 4.00 pm - Tea Break

Session VII : Chairperson - Samit Adhya4.00 - 04.50 pm

Role of Molecular Imaging in Drug Discovery Mathew ThakurTJUH, Philadelphia

04.50 – 06.00 pm - POSTER DISCUSSION06.00 – 07.00 pm Visit to CRF

07.00 – 9.00 pm - Cultural Program and Dinner (CCMB Terrace)



thWednesday, 10 November, 2010

9.30 - 11.30 am - Panel Discussion

11.30 – 12.00 Noon - Wrap up

12.00 Noon - Lunch & Disperse

Kakarla Subbarao Jyostna Rao

John DelliSanti Surender Reddy

Neeraj GuptaB.S. Dwarakanath

Indo-US Workshop on “Applications of Molecular Imaging in Health, Disease and Drug Discovery”

Organizers

Dr. Gopal PandeScientist, Centre for Cellular & Molecular BiologyUppal Road, Habsiguda, Hyderabad 500 007

Email: [email protected]

Direct Tel: +91-40-27192605, Fax: +91-40-27160311

Dr. Rao Papineni Senior Principal Investigator , Res. & Development

Carestream Health Inc, USA 4, Research Drive, WoodBridge, CT 06525

Email: [email protected] Tel 203-676-1163 (USA): +91-9177532584 (India)

LIST OF PARTICIPANTS

SPEAKERS

INVITEES

S. No.

S. No.

Name of the Speaker

Name of the Speaker

Institute & Address

Institute & Address

City

City

Email

Email

From USA

1

1

Dr. Rheal Towner

Dr. Ch. Mohan Rao

Director of Advanced Magnetic Resonance Center, 825,

thNortheast, 13 Street

Director, Centre for Cellular & Molecular Biology

Oklahoma City

Hyderabad

omrf.org

2

2

Dr. Mathew Thakur Thomas Jefferson University Hospital

CEO, Carestream Health Inc

Philadelphia mjefferson.edu

3

3

4

5

Dr. Mark Kester Distinguished Professor, Penn State University

Science Officer, Indo-US Science and Technology Forum

Hershey, PA

New Delhi

mpsu.edu

indousstf.org

Dr. Ali Arbab Associate Scientist and Director, Cellular & Molecular Imaging Laboratory, Henry Ford Hospital

Detroit shfh.edu

Dr. Kit Lam Chair, Dept. of Biochemistry and Molecular Medicine, UC Davis Cancer Centre

Davis, CA Kucdavis.edu

6 Dr. Belinda SetoDeputy Director, National Institute of Biomedical Imaging and Bioengineering

Bethesda

Indo-US Workshop on “Applications of Molecular Imaging in

Health, Disease and Drug DiscoveryCentre for Cellular & Molecular Biology, Hyderabad, India

Nov 8-10, 2010

5

ccmb.res.in

Dr. John DelliSanti WoodBridgecarestreamhealth.com

Dr. Nishritha Bopana




6

From India

7 Dr. Gopal Kundu Scientist, National Centre for Cell Science

Pune kunducom

8

9

10

11

12

13

14

15

16

Dr. P.K. GuptaHead, Laser Biomedical Applications & Instrumentation Division, CAT

Indore pkgupta

Scientist, Centre for Cellular & Molecular Biology Hyderabad abpateln

Dr. Anant Patel

Dr. R.N. Jagannathan Prof. & Head, MRI Facility, All India Institute of Medical Sciences

New Delhi

jagancom,jagancom,

nrjgj@in

Scientist, Indian Institute of Chemical Biology

Kolkata samit.res.in Dr. Samit Adhya

Dean, Amrita Institute of Medical Sciences and Research Centre, Amrita Lane AIMS Ponekkara Post

shantinair@edunaircom

Dr. Shanti K. Nair Kochi

Dr. Kakarla Subbarao

Dr. Arvind K. Chaturvedi

Dr. B Ravindran

Dr. Arun K Gupta

Ex-Director, NIMS,KREST Building, Sheikpet, Hyderabad. Tel: 23560005

Prof. & Head, Department of Radiology and Imaging, Rajiv Gandhi Cancer Institute & Research Center

Director, Institute of Life Sciences

Professor and HeadDepartment of Imaging Sciencesand Interventional Radiology, Sree Chitra Tirunal Inst. for Medical Sciences & Technology

Hyderabad

New Delhi

Bhubaneswar

Trivandrum

kakarlasubbaraocom

arvindatcom

balaraviinravindrancom

gupta@ingupta@[email protected]

1

2

3

4

5

6

PANEL DISCUSSION

Dr. Jyotsna Rao Apollo Gleneagles PET CT Centre Hyderabad jyotsnaelcom

Dr. Kakarla Subbarao Ex-Director, NIMS, Hyderabad kakarlasubbaraocom

[email protected]. John DelliSanti CEO, Carestream Health Inc.

CEO, Vijaya Dignostics

WoodBridge

Dr. Surender Reddy Hyderabad

Mr. Neeraj Gupta

Dr. B.S. Dwarakanath

ncom

bo in

Imperial Life Sciences

INMAS Delhi



7

Gurgaon

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Dr. Sujata Mohanty

Dr. Marshal

Dr. Shashi Singh

Dr. N. Madhusudhana Rao

Dr. Lekha Dinesh Kumar

Mr. G. Srinivas

All India Institute of Medical Sciences

Scientist, Centre for Cellular &Molecular Biology

Scientist, Centre for Cellular & Molecular Biology



Technical Officer, Centre for Cellular &Molecular Biology

New Delhi

Hyderabad

Hyderabad

Hyderabad

Hyderabad

Hyderabad

sujmohantyin

marshal.in

shashisin

madhuin

lekhan

gsri.in

Mr. T. AvinashrajSTA , Centre for Cellular & Molecular Biology Hyderabad taraj.in

Dr. Anil Misra Delhi akmishra.in



8

Dr. M. Ramanadham

Dr. B.S. Dwarakanath

Dr. E.J.J. Samuel

School of Life Sciences, University of Hyderabad Hyderabad mrnsl@in

Scientist, Head, Division of Radiation Biosciences, Institute of Nuclear Medicine & Allied Sciences

Scientist, Institute of Nuclear Medicine & Allied Sciences

Delhibsd.org

bsd@iin

Sr. Professor of Physics, Photonics Division, VIT University

Vellore jebamonica.in

Dr. N. Harishankar

Mr. Sachindra

Parashar

Scientist, National Institute of Nutrition Hyderabad hsnemanicom

National Centre for Cell Science Pune sachindrabiotech@

Dr. Lakshmi Chelluri Global Hospitals Hyderabad apparusucom

Dr. Aleem Ahmed Khan

Scientist, Deccan Medical College of Sciences

Hyderabad aleemrediffmail.com

PARTICIPANTS

18

19

GurgaonMr. Jai Raj Imperial Life Sciences jairajom

Ms. Supriya Saraswati National Centre for Cell Science Pune



9

16

17

Ms. Sangeeta Chakraborty

MCBL, Indian Institute of Science [email protected]

Dr. Daniela Ahl Imperial Life Sciences Gurgaon daniela..com

POSTER TITLES

1. Specificity in Gene Silencing Using Multifunctional Nanoparticles1 1 2 2Rao V. Papineni , Tao Ji , Thirupandiyur Udayakumar , Mohammed M. Shareef , Mansoor M.

2 2Ahmed , Alan Pollack1Carestream Molecular Imaging, Carestream Health, Inc., New Haven, CT, USA;2Radiation Oncology, University of Miami, Miami, FL, USA.

2. Bisphosphonate targeting to specific skeletal sites in mice with whole body vibrationRao V L Papineni. Sean Orton, Tao ji, Hans Schmitthenner, William McLaughlin, Douglas Vizard and Jingyi PanCarestream Molecular Imaging, Carestream Health Inc., New Haven, CT

3. Roadmap for molecular imaging in emerging economies-A focus on nuclear imagingJyotsna Rao and Rao V. L. Papineni,Apollo Hospitals and Carestream Health Inc.

4. Safety and efficacy of the human fetal liver derived stem cell transplantation as supportive modality in the management of end stage Decompensated liver cirrhosis – A2 year follw up studyHabibullah CM*, Aejaz Habeeb M*, Aleem A. Khan*, Mahaboob V Shaik*, Parveen N*, Rajendraprasad A*, Mohammed A Aleem*, Srinivas G†, Avinash Raj T†, Santosh K Tiwari*, Kumaresan K‡ , Venkateswarlu J*, Gopal Pande†, Linda Powers#*Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences, Kanchanbagh, Hyderabad, Andhra Pradesh, India.†Center for Cellular and Molecular Biology, Uppal Road, Hyderabad ‡KK Scan Centre, Somajiguda, Hyderabad #Vesta Therapeutics, Inc., 801 Capitola Dr., Suite 8, Durham, NC 27713, USA.

5. Techniques to Improve 3D Optical Imaging Quantification and Sensitivity Jai Raj Imperial Life Sciences P. Ltd., Gurgaon

6. Invivo imaging of murine bone marrow and human foetal liver cells in SCID & nude mice1 1 1 2 1T. Avinash Raj , G. Srinivas , J. Mahesh Kumar , Aleem A. Khan , Gopal Pande

1 Center for Cellular and Molecular Biology, Uppal Road, Hyderabad.2 Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences,

Kanchanbagh, Hyderabad.



7. Evaluation of the autologous bone marrow derived mononuclear cell therapy in the infarcted myocardium by imaging analysis – a case report

†Authors: Alla GK Gokhale., Lakshmi Kiran Chelluri., Kumaresan K. , Subramanyam G., Sudhakar K., Satish Vemuri., Tanya Debnath., Ratnakar KS

†Affiliations: Global Hospitals, Lakdi-ka-Pool, Hyderabad., KK Nuclear Clinic, Somajiguda, Hyderabad



ABSTRACTS



12

Molecular Medicine: Present and Future

Advances in molecular medicine have the potential and promise to transform medicine from the “see

and treat” paradigm to one that emphasizes personalized, predictive and preventive strategies. To

facilitate the transformation, the U.S. National Institutes of Health support and conduct research that

capitalizes on genomic and proteomic data as a basis for developing diagnostic technologies and

targeted therapy at the cellular and molecular levels. As an example, I plan to discuss a research

project that aims to identify circulating cells and molecular markers in blood samples that will

characterize subsets of patients for targeted therapies. The technology involves microfluidic chips

designed with high throughput capability to screen multiple patient blood samples to capture

signature cells with subsequent identification of the disease genomic, proteomic or molecular

signatures. The platform can be developed to integrate these tests such that a comprehensive picture

of the patient's condition is available to the physician. Immediate applications are envisioned in

cancer and potentially atheromatous disease (the two leading causes of death).

The comprehensive platform technologies may transform medicine by: targeting therapies specific

for patients; predicting responsiveness to therapy; monitoring disease progression and treatment

response at the cell/molecular level to expedite consideration of alternative therapies and probing

the fundamental biology of disease resolution or progression. This should lead to more precise

diagnoses, personalized treatment for patient sub-groups, improved outcomes and reduced cost due

to preventing advanced stages of disease.

Belinda Seto, Ph.D.Deputy Director, National Institute of Biomedical Imaging and Bioengineering

Bethesda, USA

Role of in vivo Magnetic Resonance Spectroscopic Imaging (MRSI)

and Diffusion MRI (DWI) in the Evaluation of Prostate Cancer

N. R. Jagannathan

Department of NMR and MRI Facility All India Institute of Medical Sciences

New Delhi – 110029, India.



13

Prostate cancer is the most common male cancer in the West and second most common cause of

cancer related deaths. The incidence of prostate cancer in India is however lower but a steady increase

has been observed in recent years especially in urban areas like Delhi and Mumbai. The key to

treatment is the detection of malignancy at an early stage and monitoring the tumor response to

therapy. The challenges in detection, localization and staging of prostate cancer of existing methods

have prompted us to take the investigation of the role of various MR methodologies in a large cohort

of Indian men prior to biopsy.

The identification of suspicious areas of malignancy are normally carried out using magnetic

resonance imaging (MRI), magnetic resonance spectroscopic imaging (MRSI) and diffusion weighted

imaging (DWI). Till now nearly 250 men were studied using these MR methods in our Institute. Our

data showed that the apparent diffusion coefficient may be a reliable marker to differentiate normal,

benign and malignant prostate tissues similar to the metabolite ratio obtained from MRS. Further; our

results demonstrated that the combined use of MRSI and DWI improved the diagnosis of prostate

cancer.

1The role of 3D H MRSI in directing TRUS guided biopsy of prostate was also evaluated in a large cohort

of patients. The z – and x – coordinates of the voxels suspicious of malignancy from MRSI were used to

direct TRUS guided needle biopsy. The site of biopsy was confirmed by post biopsy MRI and MRSI. MRI

showed the site of biopsy as hypo-intense area on gradient T2 weighted image (T2*) while the MR

spectrum showed reduced citrate and increased choline.

In this talk, our experience on the use of MRSI and DWI methods in the diagnosis of prostate cancer in

Indian men especially in clinically challenging cases of patients with PSA level in the range of 4 – 20

ng/mL and/or abnormal DRE will be presented.



Magnetic resonance imaging (MRI) and spectroscopy (MRS) play important roles as

diagnostic tools for many cancers, including assessments on tumor growth, angiogenesis,

bio-physical parameters, such as diffusion and perfusion, and response to therapy.

Molecular MRI (mMRI) has made some important contributions in the study of angiogenic,

inflammation and apoptotic markers in cancer. A brief discussion on advances in the

development of mMRI contrast agents will be presented. Examples on the use of high

resolution MRI and MRS at 7.0 Tesla to obtain molecular imaging information on

angiogenesis, inflammation and tumor metabolism in rodent pre-clinical models for gliomas

and hepatocellular carcinomas (HCC) will be presented. The concept of mMRI involves the

coupling of an affinity molecule that recognizes a specific target marker (e.g. vascular

endothelium growth factor receptor 2 (VEGF-R2), c-Met, or inducible nitric oxide synthase

(iNOS)) and MRI contrast signaling agents such as gadolinium-ligand- or iron oxide

nanoparticle (IONP)-based constructs. Chemical shift imaging (CSI) as a method to assess

lipid metabolism in tumor models will also be presented. MR methods offer the unique

opportunity to be used as tools to obtain non-invasive in vivo morphological/anatomical,

vascular, metabolic, functional and molecular information in rodent pre-clinical models in

cancer.

1 1 1 1 1 1Rheal A. Towner , Nataliya Smith , Ting He , Sabrina Doblas , Philippe Garteiser , Debra Saunders , 2 2Robert Silasi-Mansat and Florea Lupu

1 2Advanced Magnetic Resonance Center, and Cardiovascular Biology,Oklahoma Medical Research Foundation

th825 N.E. 13 St., Oklahoma City, OK 73104 U.S.A.Phone: (405) 271-7383

FAX: (405) 271-7254Email: [email protected]

Pre-Clinical Molecular Imaging Applications in Cancer Research


P.K. GuptaCAT, Indore Laser Biomedical Applications and Instrumentation Division,

Raja Ramanna Centre for Advanced Technology,

Indore – 452013, India.

E-mail: [email protected]

Use Of Optical Spectroscopic And Imaging Techniquesfor Biomedical Diagnosis

I shall provide an overview of the studies carried out at Laser Biomedical Applications and

Instrumentation Division, RRCAT on the development and utilization of optical techniques for

biomedical imaging and diagnosis. First I shall discuss some representative biomedical imaging

studies carried out using the optical coherence tomography set ups developed at RRCAT. Work

carried out at RRCAT on the use of optical spectroscopy for biomedical diagnosis will be discussed

next and the results of our recent study on a comparative evaluation of the performance of laser

induced fluorescence spectroscopy and Raman spectroscopy for in-vivo diagnosis of the cancer of

oral cavity will be presented. Finally I shall also describe some aspects of the Raman spectroscopic

studies carried out by us on single, optically trapped Red Blood Cells obtained from blood samples

from healthy volunteers and from patients suffering from malaria.




Image guided Interventions

A.K. Gupta

Abstract not received

Professor and HeadDepartment of Imaging Sciences

and Interventional Radiology, Sree Chitra Tirunal Inst. for Medical Sciences & Technology

Thiruvananthapuram

The authors discuss new methodologies and treatments of cancer using molecular imaging and

targeted therapeutics using non-toxic nanoparticles. At the Amrita Centre for Nanosciences a range

of multifunctional nanoparticles have been developed with combined fluorescence, X-Ray and MRI

contrast capability. When such particles can be targeted to cancer cells there is the possibility of early

detection by both anatomical and molecular imaging modalities including the opportunity of

delivering drugs directly to the cells in question. Efforts are currently under way to target cancer stem

cells in addition to cancer cells to enhance early detection and therapeutic efficiencies. The results of

some of our current studies will be presented at this workshop.



17

Combined Anatomical and Molecular Imaging with Therapeutics using Biocompatible Nanoparticles

Shantikumar Nair and K.ManzoorAmrita Centre for Nanosciences

Amrita Vishwa Vidyapeetham, Kochi



18

From combinatorial chemistry to nanoparticles to cancer imaging and therapy

1 1 1 1 1 1 1Juntao Luo , Kai Xiao , Yuan-pei Li , Joyce Lee , Nianhuan Yao , Wenwu Xiao , Yan Wang , Michael 2, 3 4 1Kent Holland Cheng , Gangyu Liu , and Kit S. Lam *

1 Department of Biochemistry & Molecular Medicine,Division of Hematology & Oncology, UC Davis 2 3 4Cancer Center, Veterinarian School of Medicine, College of Biological Sciences, Department of

Chemistry, University of California Davis, CaliforniaCorresponding author ([email protected])

Recently we have developed a novel amphiphilic polymer (telodendrimer), comprised of linear

polyethylene glycol, lysine and cholic acid, that can self-assemble under aqueous condition to form

stable micelles. The telodendrimer is prepared by Fmoc-chemistry with standard peptide synthesis

methods. Various hydrophobic drugs such as paclitaxel, etoposide, doxorubicin, SN-38, vinblastine,

vincristine, temsirolimus, and bortezomib can be easily incorporated into the nanoparticles with high

loading capacity. This nanoplatform is highly versatile. We can readily tune the particle size by varying

the configuration and composition of the telodendrimer. DiD near infra-red hydrophobic dye can be

used as drug surrogate for loading inside the nanoparticles for in vivo biodistribution studies in nude

mice bearing ovarian cancer xenograft. Particles smaller than 60 nm were found to preferentially

target the tumors, whereas particles at 150nm were found to concentrate at the liver and lung. The

surface chemistry of the nanoparticles can be easily modified by derivatizing the distal polyethylene

glycol tail of the telodendrimers with short oligopeptides comprised of D-lysine or D-aspartate.

Undesirable liver uptake was found to be very high for those nanoparticles with high surface charges,

either positive or negative. In contrast, liver uptake was very low but tumor uptake was very high

when the surface charge was slightly negative, e.g. one aspartate at the PEG terminus of each

telodendrimer. In the last few years, we have use one-bead one-compound combinatorial library

method to identify tumor targeting ligands. Ligation of one of these ligands OA02, that targets αvβ3

integrin, to the nanoparticle surface further improved tumor targeting and intracellular uptake at the

tumor site. In addition to xenograft models, we have also demonstrated that the nanoparticles could

preferentially target spontaneous mammary tumors of PyV-MT transgenic mice. We have just

completed a Phase I study of placiltaxel-loaded nanoparticles in companion dogs with lymphoma and

showed that the drug is safe. A Phase II study of the same preparation in companion dogs with solid

tumor is on-going.

As a part of our ongoing project on RNA therapeutics of mitochondrial disease, we have developed

methods to visualize fluorescent-tagged RNA in living cells and in live animals. The RNA is

prepared by incorporation of Alexa Fluor (AF)-or BODIPY-Texas Red (TR) tagged ribo UTP during in

vitro RNA directed transcription of DNA templates. The corresponding fluorophores can be

excited by 488 and 633 nm lasers respectively. RNA labeled with either fluorophore was taken up

by cultured cells in presence of a protein carrier. Live cell confocal microscopy of cells doubly

labeled with RNA and green (GFP) or red (RFP) fluorescent protein enabled the imaging of specific

steps of the intracellular transport process of the RNA in real time. In addition, we could visualize

its fate in mice after local or systemic administration of BODIPY-TR labeled RNA by live animal

imaging. These developments will facilitate the study of the transport and metabolism of

biologically active RNA in vitro and vivo.



Imaging RNA in cells and in animals

Sukanta Jash, Biraj Mahato and Samit Adhya

Division of Molecular and Human Genetics, Indian Institute of Chemical Biology,

4 Raja S. C. Mullick Road, Calcutta 700032, India.



Immunosuppression is a major obstacle to effective treatment of cancer, and can be a contributing

factor to therapy resistance. Several immunosuppressive cells have been identified, including

myeloid-derived suppressor cells (MDSC), and these have been shown to be increased in laboratory

models of cancer as well as cancer patients. MDSCs directly interfere with T cell mediated immunity,

and dendritic and natural killer cell function. Therefore significant effort is underway toward the

development of therapies that decrease MDSCs. The current study demonstrated that a previously

described deep tissue imaging modality, which utilized encapsulation of indocyanine green within a

calcium phosphosilicate-matrix nanoparticle ((ICG-CPSNP), can be utilized as an immunoregulatory

therapeutic agent. The theranostic application of ICG-CPSNPs as a photosensitizer for photodynamic

therapy (PDT) was shown to effectively block tumor growth in murine breast cancer models by

decreasing MDSCs while increasing immune effectors. As phosphorylated sphingolipid metabolites

have been shown to have immunomodulatory roles, it was hypothesized that the reduction of MDSCs

by ICG-CPSNP PDT was dependent upon bioactive sphingolipids. ICG-CPSNP PDT induced a

sphingosine kinase-dependent increase in dihydrosphingosine-1-phosphate (dhS1P). Indeed,

isolated MDSCs were decreased by treatment with dhS1P, but not sphingosine-1-phosphate, while

dhS1P induced a concomitant expansion of antitumor B cells. Adoptive transfer of these dhS1P-

induced B cells into tumor-bearing mice effectively blocked breast cancer tumor growth. Collectively,

these findings revealed that PDT utilizing the theranostic agent ICG-CPSNP, also behaved as a photo-

immunotherapy in breast cancer by prompting a decrease in immunosuppressive MDSCs and an

increase in immune effectors.

Conflict of Interest Disclaimer: Penn State Research Foundation has licensed calcium phosphosilicate

nanotechnology to Keystone Nano, Inc. (State College, PA). J.A. and M.K. are CSO and CMO of

Keystone Nano, respectively.

Nano"solutions" as Theranostics Dihydrosphingosine-1-Phosphate Triggers Expansion of Antitumor Immune Effectors

During Photoimmuno-Nanotherapy

Brian M. Barth, James M. Kaiser, Sriram S. Shanmugavelandy, Todd E. Fox, Erhan İ. Altinoğlu, Sarah A. Knupp. Georgina V. Bixler, Robert M. Brucklachen, Willard M. Freeman, Timothy M. Ritty, James

,H. Adair, and Mark Kester

The Pennsylvania State University College of Medicine, Hershey, PA 17033, USA



The molecular imaging of tumor growth, angiogenesis and metastasis recently represent a major area

of research for the diagnosis and targeted therapy of many solid tumors in the area of cancer biology.

An

optimized set of high efficiency filters and spectral un-mixing algorithms allow taking full advantage of

bioluminescent and fluorescent reporters across the blue to near infrared wavelength region. In our

laboratory, we have recently developed many tumors in mice models and the effects of many drugs

have been tested and subsequently these tumors have been analyzed based on bioluminescence

technology. The analysis of some of these tumor images will be demonstrated during this workshop.

In vivo bioluminescent and fluorescent imaging allows the non-invasive detection and quantification

of orthotropic, metastatic and spontaneous tumors in the whole body of mouse. Many imaging

technology has been optimized for high sensitivity, such that micro metastases can be detected.

In Vivo Imaging of Tumors in Mice using Bioluminescence and Fluorescence Technologies

Gopal Kundu

Scientist-F, National Centre for Cell Science, Pune 411 007, India;

E-mail:[email protected]

Personalized treatment using stem, modified or genetically engineered, cells is becoming a reality in

the field of medicine, in which allogeneic or autologous cells can be used for treatment and possibly

for early diagnosis of diseases. Hematopoietic, stromal and organ specific stem cells are under

evaluation for cell-based therapies for cardiac, neurological, autoimmune and other disorders.

Cytotoxic or genetically altered T-cells are under clinical trial for the treatment of hematopoietic or

other malignant diseases. Before using stem cells in clinical trials, translational research in

experimental animal models are essential, with a critical emphasis on developing noninvasive

methods for tracking the temporal and spatial homing of these cells to target tissues. Moreover, it is

necessary to determine the transplanted cells, engraftment efficiency and functional capability.

Tracking of migration and homing of systemic or locally administered cells using MRI and other

complementary imaging modalities have passed the infancy. Now the technology is in a state where

robust experiments can be designed in translational studies utilizing cellular magnetic resonance

imaging (CMRI), nuclear medicine techniques and optical imaging. Tagging cells with reporter genes,

fluorescent dyes or different contrast agents transforms them into cellular probes or imaging agents.

In this presentation we will discuss the methods to transform cells into probes for in vivo imaging,

along with their advantages and disadvantages as well as the future clinical applicability of cellular

imaging method and corresponding imaging modality.

Points:1) Potential application of cells as diagnostic and therapeutic probes.2) In vitro making of potential therapeutic CTLs for malignant diseases3) Use of stem cells as gene carrier and delivery vehicles4) Use of multimodality approaches

In Vivo Cellular Imaging and its Role in Cell Therapy

Ali Arbab, MD, PhDAssociate Scientist

Cellular and Molecular Imaging Laboratory, Department of Radiology, Henry Ford Hospital, Detroit, MI, USA

&Associate Professor

Department of RadiologyWayne State University School of Medicine, Detroit, MI, USA



22



Glutamate and GABA are the major excitatory and inhibitory neurotransmitters in the matured 1mammalian central nervous system and together account for ~90% neurotransmitter pool in cortex .

These neurotransmitters play major roles in energy metabolism, cortical excitability, and cognitive 13function. It is well established that neuronal astrocytic substrate cycle exists in the brain. In vivo C

NMR studies have established that glutamatergic energetics is supported by oxidative glucose 1,2metabolism . Further, GABA contributes significantly to total neurotransmission and glucose

3oxidation by neurons in normal brain . These findings have led to specific predictions about the 4,5fundamental role of glucose in the support of neurotransmitter cycling . Dysfunction in glutamate

and GABA pathways are associated with many neurological and neuropsychiatric disorders. Thus,

glutamate, GABA and glutamine metabolism is of major importance to brain function and cerebral

well-being.

13In this presentation, I will discuss the strategy involving infusion of C labeled substrates 13together with C NMR spectroscopy to measure fluxes through glutamatergic and GABAergic

pathways. We are extending these measurements in mice to understand cerebral metabolism in

different cerebral disorders. I will be presenting our recent data for the investigation of flux through

glutamatergic and GABAergic pathways in healthy as well as in various abnormal/diseased conditions

such as neurodegeneration (Parkinson's, Alzheimer's), addiction (nicotine, alcohol).

References: 1. Sibson et al (1998) Proc Natl Acad Sci 95:316. 2. Patel et al (2004) J Cereb Blood Flow

Metab 24:972. 3. Patel et al (2005) Proc Natl Acad Sci 102:5588. 4. Magistratti et al (1998) Science

283:5801. (3) Hyder et al (2005) J Cereb Blod Flow Metab 26:865.

13 CNMR Investigations of Excitatory and Inhibitory Neurotransmission in Cerebral Disorders

Anant B. PatelNMR Microimaging & Spectroscopy,

Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, India



Despite the great strides made in drug discoveries, many diseases have continued to take toll on

human lives and its quality. Recent advances in molecular biology and genomics have resulted into an

increasing number of novel biomarkers. These biomarkers can contribute extensively into the

development of translational medicine to diagnose diseases as well into the design of personalized

medicine to treat diseases. In the development of such drugs, molecular imaging can play a pivotal

role.

Cancer for example, thought to be initiated by genomic modulation at a cellular level, is known to

overexpress key proteins and RNAs of specific signaling pathways. These have become a hallmark of

many cancers, and an array of such biomarkers have been identified. Similarly the resolution of

common infection, in part, is associated with neutrophil apoptosis accelerated by reduction of

prosurvival factors such as lipopolysaccharides (LPS) and certain cytokines such as IL-1B. Apoptotic

neutrophils are presented in the outer layers, with Phosphatidylserine (PS), which is ordinarily

sequestered in the plasma membrane inner leaflet, and provides itself as an excellent target for

localizing infectious foci in vivo, and for monitoring the effectiveness of its treatment.

We hypothesize that targeting such biomarkers with specific fluorophores for optical imaging or with

radiolabeled specific biomolecules such as peptides on peptide nucleic acids for PET or SPECT imaging

of malignant lesions can help to address the challenges of drug discovery.

Examples of investigations from this laboratory will illustrate approaches not only for diagnosis of

diseases but also for their treatment, stratification and to determine the effectiveness of their

therapeutic interventions.

Supported by NIH CA 10923, BB 001809, ISIORR23709, PA ME-03-184, DOE/BER 63055, NIH CO27175,

NIH CA148565 and NuView, Inc.

Mathew L. Thakur, PhD

Department of Radiology, Thomas Jefferson University, Philadelphia, USA

ROLE OF MOLECULAR IMAGING IN DRUG DISCOVERY

first indo-us bilateral meet on molecular imaging

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