Identification of lymph node (LN) metastasis is essential for staging of solid tumors, and as a result, surgeons focus on harvesting significant numbers of LNs during ablative procedures for pathological evaluation. Isolating those LNs most likely to harbor metastatic disease can allow for a more rigorous evaluation of fewer LNs. Here we evaluate the impact of a systemically injected, near-infrared fluorescently-labeled, tumor-targeting contrast agent, panitumumab-IRDye800CW, to facilitate the identification of metastatic LNs in the ex vivo setting for head and neck cancer patients. Molecular imaging demonstrates a significantly higher mean fluorescence signal in metastatic LNs compared to benign LNs in head and neck cancer patients undergoing an elective neck dissection. Molecular imaging to preselect at-risk LNs may thus allow a more rigorous examination of LNs and subsequently lead to improved prognostication than regular neck dissection.

The MYC oncogene drives T-and B-lymphoid malignancies, including Burkitt's lymphoma (BL) and Acute Lymphoblastic Leukemia (ALL). Here, we demonstrate a systemic reduction in natural killer (NK) cell numbers in SRα-tTA/Tet-O-MYC ON mice bearing MYC-driven Tlymphomas. Residual mNK cells in spleens of MYC ON T-lymphoma-bearing mice exhibit perturbations in the terminal NK effector differentiation pathway. Lymphoma-intrinsic MYC arrests NK maturation by transcriptionally repressing STAT1/2 and secretion of Type I Interferons (IFNs). Treating T-lymphoma-bearing mice with Type I IFN improves survival by rescuing NK cell maturation. Adoptive transfer of mature NK cells is sufficient to delay both T-lymphoma growth and recurrence post MYC inactivation. In MYC-driven BL patients, low expression of both STAT1 and STAT2 correlates significantly with the absence of activated NK cells and predicts unfavorable clinical outcomes. Our studies thus provide a rationale for developing NK cell-based therapies to effectively treat MYC-driven lymphomas in the future.

34The MYC oncogene drives T and B lymphoid malignancies, including Burkitt's lymphoma (BL) 35and Acute Lymphoblastic Leukemia (ALL). Using CyTOF, we demonstrate a systemic reduction 36 in natural killer (NK) cell-mediated surveillance in SRα-tTA/Tet-O-MYC ON mice bearing MYC-37driven T-lymphomas, due to an arrest in NK cell maturation. Inactivation of lymphoma-intrinsic 38 MYC releases the brakes on NK maturation restoring NK homeostasis. Lymphoma-intrinsic 39 MYC arrests NK maturation by transcriptionally repressing STAT1/2 and secretion of Type I 40 Interferons (IFNs). Treating T-lymphoma-bearing mice with Type I IFN improves survival by 41 rescuing NK cell maturation. In MYC-driven BL patients, low expression of both STAT1 and 42 STAT2 correlates significantly with the absence of activated NK cells and predicts unfavorable 43 clinical outcomes. Adoptive transfer of mature NK cells is sufficient to delay both T-lymphoma 44 growth and recurrence post MYC inactivation. Our studies thus provide a rationale for 45 developing NK cell-based therapies to effectively treat MYC-driven lymphomas in the future. 46Oncogene addiction is the phenomenon where a cancer becomes causally dependent on a single 47 'driver' oncogene for the maintenance of its proliferation and survival 1,2 . Inhibition of the driver 48 oncogene forms the basis of oncogene targeted therapy 1-3 . Hence, a potential Achilles' heel of 49 human malignancies lies in their addiction to oncogenes such as MYC, RAS and BCR-ABL1 4,5 . 50Targeted therapy has been successful against BCR-ABL1 (Philadelphia/Ph + )-induced leukemia 51 6,7 , while therapies that successfully target MYC or RAS remain to be developed. 52In order to develop targeted therapies for MYC-driven cancers, it is vital to understand 53 whether MYC regulates both cell autonomous and non-autonomous processes, including host 54 immunity. Oncogene addiction was previously assumed to be cell autonomous and immune-55 independent 8 . Many recent studies illustrate that MYC and other oncogenes alone or 56 cooperatively may regulate the tumor microenvironment and host immune responses in multiple 57 tumor types 9-16 . In particular, two reports highlighted that oncogenes may cooperate to more 58 globally regulate the host immune system 12,16 . 59We have used a particularly tractable approach for studying the role of the host immune 60 system during MYC-driven tumorigenesis through tetracycline (tet)-system regulated transgenic 61 mouse models of cancer. Our inducible mouse models enable us to study how MYC inactivation 62 elicits tumor regression through both cancer-intrinsic and cancer-extrinsic host immune-63 dependent mechanisms 11,13 . CD4 + T cells appear to be essential for the sustained regression of 64 cancers upon MYC inactivation 11,13 . However, the changes in immune landscape during primary 65 MYC-induced tumorigenesis in these models remain to be delineated. 66Our goal was to delineate the global immunological changes resulting from primary overt 67 MYC-driven lymphomagenesis, to identify...

Lung cancer is the leading cause of cancer-related deaths worldwide. Most patients present with advanced inoperable disease. Traditionally, responses to treatments are evaluated using different imaging modalities, which can sometimes be confusing. This is particularly more relevant in stage 3 disease where, after radiation therapy, persistent tumors on scans can represent active disease or scar tissue. We have been evaluating role of circulating tumor cells (CTCs) in that setting. Here we present the case of a 68-year-old male with stage 3 disease whose primary tumor responded to chemoradiotherapy on imaging, but whose CTC count was higher than the pre-treatment value. The patient later developed liver metastases. In this case, the CTC count more accurately predicted the patient's prognosis and highlights the need for exploration of the CTC count as a tool supplemental to imaging modalities.

Circulating tumor cells (CTCs) are indicative of metastatic disease in multiple types of solid tumors. Technologic advances in CTC enrichment have yielded profound variability in both quantity and phenotypic characteristics of CTCs. While size-based exclusion methods have improved the sensitivity of CTC capture, their diminished specificity requires subsequent robust cytopathologic identification of CTCs. In this study, we compared CTC counts from Isolation by Size of Epithelial/Trophoblastic Tumor cells (ISET®) filters sequentially stained by May-Grünwald/Giemsa (MGG), immunocytochemistry (ICC)/hematoxylin, and ICC/hematoxylin/eosin, followed by corresponding CTC criteria. An immune and endothelial cell cocktail of CD45/CD11b/CD31 antibodies adequately ruled out immune and endothelial cells, yet a substantial number of atypical morphologies with nuclear irregularity (i.e., circulating non-hematological/endothelial cells; CNHCs) were detected in both breast cancer cases and non-cancerous controls following hematoxylin nuclear counterstain. Cytoplasmic staining with eosin, significantly diminished CNHC counts. In conclusion, detection of CTCs from ISET filters using chromogenic ICC is feasible in conjunction with identification criteria of nuclear irregularity, negative reactivity to immune and endothelial cell markers, and presentation of visible cytoplasm.