The broad goal of this proposal is to use fast in vivo click chemistries to enable biomedical imaging with affinity ligands such as monoclonal antibodies. The candidate, Neal Devaraj, has received extensive training in the field of chemistry and now wishes to transition to biomedical research. His immediate goal is to further develop skill sets through immersion in translational research and to make significant contributions to the field of in vivo imaging. Long term, the candidate wishes to make a full transition from chemist to independent biomedical research scientist. The candidate proposes to carry out the proposal at the Harvard/MGH Center for Systems Biology under the mentorship of Dr. Ralph Weissleder MD PhD. In addition to the research proposal, his career development plan includes supplemental coursework, mentored readings, journal clubs, attendance and presentation at several seminars and conferences, practical exposure to the imaging sciences, and close formal interaction with both his mentor and an advisory committee consisting of Prof. Robert Langer, Prof. Moungi Bawendi, and Prof. Marcelo Di Carli. The research plan aims to develop platform chemistries to allow site specific in vivo targeting of affinity ligands once they have homed to their target (i.e. decoupled administration of affinity ligand and imaging reporter). Though a large number of targeting ligands have been developed to date, target-reporter constructs have inherent pharmacokinetics which are undesirable for in vivo imaging. To achieve site specific coupling, the candidate proposes to work with the recently introduced tetrazine/strained dienophile cycloaddition. This extremely rapid reaction is highly selective, irreversible, and can be performed in physiological conditions. The proposal aims to build on this cutting-edge technology and to develop generic in vivo click reactions for molecular imaging with positron emission tomography (PET).
The specific aims are to 1) develop and characterize a panel of fast "click" reactions for radioisotope labeling and 2) pursue in vivo click labeling for biomedical imaging. To test this method, the candidate will create dienophile affinity ligands against A33 glycoprotein, a surface persistent marker that is highly expressed in human colon cancers. Mice bearing primary and metastatic colon cancer will be injected with dienophile affinity ligands. After several days of clearance, a small, readily cleared tetrazine imaging probe will be administered. Imaging with PET will be used to quantify probe localization and the method will be optimized and compared to other methods such as direct labeling of affinity ligands and the metabolic reporter 18fluorodeoxglucose. If successful, the development of in vivo "click" chemistry for biomedical imaging will provide a nearly universal methodology for in vivo imaging of virtually any targeting affinity ligand.

Public Health Relevance

This project aims to develop improved methods for biomedical imaging. If successful this would lead to detection of disease much earlier when they are curable, improved diagnosis of diseased condition so appropriate treatment can be administered, and better tools for determining the effectiveness of new therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01EB010078-06
Application #
8660300
Study Section
Special Emphasis Panel (ZEB1-OSR-B (O2))
Program Officer
Erim, Zeynep
Project Start
2010-05-15
Project End
2015-04-30
Budget Start
2014-05-01
Budget End
2015-04-30
Support Year
6
Fiscal Year
2014
Total Cost
$167,297
Indirect Cost
$12,392
Name
University of California San Diego
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Yang, Jun; Liang, Yong; Še?kut?, Jolita et al. (2014) Synthesis and reactivity comparisons of 1-methyl-3-substituted cyclopropene mini-tags for tetrazine bioorthogonal reactions. Chemistry 20:3365-75
Nichols, Brandon; Qin, Zhengtao; Yang, Jun et al. (2014) 68Ga chelating bioorthogonal tetrazine polymers for the multistep labeling of cancer biomarkers. Chem Commun (Camb) 50:5215-7
Wu, Haoxing; Yang, Jun; Še?kut?, Jolita et al. (2014) In situ synthesis of alkenyl tetrazines for highly fluorogenic bioorthogonal live-cell imaging probes. Angew Chem Int Ed Engl 53:5805-9
Seckute, Jolita; Yang, Jun; Devaraj, Neal K (2013) Rapid oligonucleotide-templated fluorogenic tetrazine ligations. Nucleic Acids Res 41:e148
Seckute, Jolita; Devaraj, Neal K (2013) Expanding room for tetrazine ligations in the in vivo chemistry toolbox. Curr Opin Chem Biol 17:761-7
Cole, Christian M; Yang, Jun; Seckute, Jolita et al. (2013) Fluorescent live-cell imaging of metabolically incorporated unnatural cyclopropene-mannosamine derivatives. Chembiochem 14:205-8
Yang, Jun; Karver, Mark R; Li, Weilong et al. (2012) Metal-catalyzed one-pot synthesis of tetrazines directly from aliphatic nitriles and hydrazine. Angew Chem Int Ed Engl 51:5222-5
Devaraj, Neal K; Thurber, Greg M; Keliher, Edmund J et al. (2012) Reactive polymer enables efficient in vivo bioorthogonal chemistry. Proc Natl Acad Sci U S A 109:4762-7
Budin, Itay; Devaraj, Neal K (2012) Membrane assembly driven by a biomimetic coupling reaction. J Am Chem Soc 134:751-3
Haun, Jered B; Devaraj, Neal K; Marinelli, Brett S et al. (2011) Probing intracellular biomarkers and mediators of cell activation using nanosensors and bioorthogonal chemistry. ACS Nano 5:3204-13

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