Tumors are comprised of a variety of cell types in addition to the malignant cells, including stroma, vasculature, and infiltrating immune cells. An understanding of the identities, numbers, and states of these other cellular components is clearly important to understanding the biology of tumors, and their responses to therapy, as outlined in the NCI Provocative Question C4. Recently, immune-modulatory therapies and adoptive cell therapies have become established as powerful therapeutic approaches in cancer, raising a critical need to be able to monitor immune responses directly within tumors. Positron emission tomography (PET) provides highly sensitive, non-invasive and quantitative imaging, which can be combined with antibody-based probes to provide molecular imaging of tissues and cells in vivo based on cell surface phenotype. Furthermore, engineered antibody fragments (such as cys-diabodies;covalent dimers of single-chain antibody fragments) have been generated with optimal kinetics enabling rapid in vivo targeting and clearance for imaging applications by immunoPET. In the proposed work, immunoPET using engineered antibody fragments that recognize CD8, a classic cell surface marker on cytotoxic T cells, will be developed to address the challenge of direct imaging of immune cell subsets in tumors.
Specific Aim 1 will focus on the need for imaging agents to be biologically inert, and will assess biological effects (activation, cytokine release) of treating T cells with anti- CD8 cys-diabodies,in vitro and in vivo.
Specific Aim 2 will employ radiolabeled anti-CD8 cys-diabodies for immunoPET assessment of immune activation in normal mice and tumor-bearing mice treated with immune stimulants such as anti-CD137 antibodies.
Specific Aim 3 will focus on immunoPET detection and quantification of CD8+T cells in tumor-bearing mice treated with antibodies that reverse immunosuppression (such as anti-PD-1 and anti-CTLA-4) and induce tumor infiltration by cytotoxic T cells. These studies will validate the role of immunoPET probes for CD8+ T cells to study responses to cancer immunotherapy in vivo, provide a deeper understanding of immune responses in living organisms, and will open the door to broad methods for imaging and quantitating immune cell subsets in preclinical models and eventually humans.
One of the great challenges in cancer biology is to understand the presence and function of the many cell types found in tumors including cells from the host's immune response. This project will develop novel imaging agents for detection of key immune cells such as CD8-expressing cytotoxic T cells, using engineered antibodies that recognize the CD8 marker. These will be combined with Positron emission tomography (PET) to provide a highly sensitive, non-invasive and quantitative approach for directly assessing immune responses, especially important for the development and evaluation of new cancer immunotherapies.
|Lu, Jianqin; Liu, Xiangsheng; Liao, Yu-Pei et al. (2017) Nano-enabled pancreas cancer immunotherapy using immunogenic cell death and reversing immunosuppression. Nat Commun 8:1811|
|Freise, Amanda C; Zettlitz, Kirstin A; Salazar, Felix B et al. (2017) ImmunoPET Imaging of Murine CD4+ T Cells Using Anti-CD4 Cys-Diabody: Effects of Protein Dose on T Cell Function and Imaging. Mol Imaging Biol 19:599-609|
|McCracken, M N; Tavaré, R; Witte, O N et al. (2016) Advances in PET Detection of the Antitumor T Cell Response. Adv Immunol 131:187-231|
|Tavaré, Richard; Escuin-Ordinas, Helena; Mok, Stephen et al. (2016) An Effective Immuno-PET Imaging Method to Monitor CD8-Dependent Responses to Immunotherapy. Cancer Res 76:73-82|
|Freise, Amanda C; Wu, Anna M (2015) In vivo imaging with antibodies and engineered fragments. Mol Immunol 67:142-52|
|Tavaré, Richard; McCracken, Melissa N; Zettlitz, Kirstin A et al. (2015) Immuno-PET of Murine T Cell Reconstitution Postadoptive Stem Cell Transplantation Using Anti-CD4 and Anti-CD8 Cys-Diabodies. J Nucl Med 56:1258-64|
|Tavaré, Richard; McCracken, Melissa N; Zettlitz, Kirstin A et al. (2014) Engineered antibody fragments for immuno-PET imaging of endogenous CD8+ T cells in vivo. Proc Natl Acad Sci U S A 111:1108-13|