Hematogenous Metastatic Spread Research Articles

The Heparan Sulfate Proteoglycan Syndecan-1 Triggers Breast Cancer Cell-Induced Coagulability by Induced Expression of Tissue Factor.

Syndecan-1 (Sdc-1) upregulation is associated with poor prognosis in breast cancer. Sdc-1 knockdown results in reduced angiogenesis and the dysregulation of tissue factor (TF) pathway constituents. Here, we evaluate the regulatory mechanisms and functional consequences of the Sdc-1/TF-axis using Sdc-1 knockdown and overexpression approaches in MCF-7 and MDA-MB-231 breast cancer cells. Gene expression was analyzed by means of qPCR. Thrombin generation and cell migration were detected. Cell-cycle progression and apoptosis were investigated using flow cytometry. In MDA-MB-231 cells, IL6, IL8, VEGF, and IGFR-dependent signaling affected TF pathway expression depending on Sdc-1. Notably, Sdc-1 depletion and TF pathway inhibitor (TFPI) synergistically affected PTEN, MAPK, and STAT3 signaling. At the functional level, the antiproliferative and pro-apoptotic effects of TFPI depended on Sdc-1, whereas Sdc-1's modulation of cell motility was not affected by TFPI. Sdc-1 overexpression in MCF-7 and MDA-MB-231 cells led to increased TF expression, inducing a procoagulative phenotype, as indicated by the activation of human platelets and increased thrombin formation. A novel understanding of the functional interplay between Sdc-1 and the TF pathway may be compatible with the classical co-receptor role of Sdc-1 in cytokine signaling. This opens up the possibility of a new functional understanding, with Sdc-1 fostering coagulation and platelet communication as the key to the hematogenous metastatic spread of breast cancer cells.

The Antibacterial Effect of Boron Compounds and Evaluation of the Effects on Biofilm Formation in the Infection Model of Klebsiella pneumoniae on the HepG2 Cell Line

Aim: Klebsiella pneumoniae causes hospital-acquired pneumonia, urinary tract infection and bacteremia in immunocompromised patients. Klebsiella pneumoniae, which has become more common recently, causes antibiotic resistance as well as pyogenic liver abscesses and hematogenous metastatic spread in humans. Developing antibiotic resistance complicates the treatment of liver infections. In our study, we aimed to evaluate the effect of boron compounds in an infection model created by Klebsiella pneumoniae 700603. Materials and Methods: Minimum inhibitory concentration and fractional inhibitory concentration studies, resistance gene levels, and HepG2 cell analyses were performed and evaluated. Results: We determined the low and high minimum inhibitory concentration values of boron components, sodium perborate monohydrate and etidote, respectively. In addition, sodium perborate monohydrate is also effective on biofilm formation. Our findings have shown that boron compounds are more effective when used in a combination. In the toxicity model created in the cellular study, the boron compounds cytotoxic effect decreased due to their antibacterial effects. Conclusion: İt seems that boron compounds are effective, and the positive effect increases when used together.

Role of Radiation Therapy in Retroperitoneal Sarcoma.

Retroperitoneal sarcoma comprises a small subset of all soft tissue sarcoma and includes various histopathologic subtypes, each with unique patterns of behavior and differential risks for local recurrence and hematogenous metastatic spread. The primary treatment modality is surgery, although even with complete macroscopic resection, recurrence is common. The rationale for the addition of radiotherapy to resection is to improve local control; however, the use of radiation therapy for retroperitoneal sarcoma is controversial, and existing data are suboptimal to guide management. Treatment decisions should be determined with multidisciplinary input and shared decision-making. When used in selected patients, radiation therapy should be delivered preoperatively; postoperative treatment is not recommended.

Leiomyosarcoma metastasis tumor in calvaria

The incidence of a metastatic tumor of the calvaria is uncommon, but it has a considerable impact in the field of neurosurgery. The hematogenous metastatic spread was common in leiomyosarcoma. Calvaria is one of the most uncommon metastatic locations for this type of cancer. A 70-year-old man with a history of rhabdomyosarcoma and thyroid cancer came with a parietal mass that rapidly enlarged, followed by sudden hemiparesis and severe cognitive impairment. Head CT scan revealed a destructive soft tissue mass, indicating a metastatic development and mass effect of tumor pushing the brain parenchymal with slight compression of the right lateral ventricle and midline deviation of 8 mm to the left side. Perioperative embolization was done, and the tumor was effectively excised through extensive excision. Histopathology indicated a malignant spindle cell mesenchymal tumor, indicating leiomyosarcoma. After the operation, the patient's neurological status was stable, and there was no sign of tumor recurrence. Recommendation for managing metastatic calvaria tumor based on histopathological findings is insufficient. The information in this case report is essential for scholarly purposes and the participant gave written informed consent for publication. For the treatment of metastases in the skull, complete excision of osteolytic calvaria lesions is indicated.

EMT-related genes are unlikely to be involved in extracapsular growth of lymph node metastases in gastric cancer

BackgroundIn gastric cancer (GC), extracapsular growth (ECG) pattern of lymph node metastases is associated with decreased overall survival rates compared to intracapsular lymph node metastases (ICG). Epithelial-to-mesenchymal transition (EMT) plays a pivotal role in hematogenous metastatic spread. Aim of the present study was to analyze if EMT related genes are involved in the growth pattern of lymph node metastases in GC. MethodsOut of our prospective database with 529 patients who underwent surgical resection for GC between 2002 and 2014 forty lymph node positive patients were identified (20 ECG, 20 ICG). The expression of 84 EMT-associated genes were analyzed by RT2 Profiler PCR Array (n = 20). Results were validated by Real-Time PCR (n = 20). ResultsGC with ECG showed differently expressed EMT related genes. GC leading to ECG showed an upregulation of three and downregulation of eleven genes. Those differences, however, could not be confirmed in PCR analysis. ConclusionsThis study demonstrates that EMT related genes are not responsible for the different growth patterns of lymph node metastases in GC. Further studies are required to evaluate the underlying mechanisms of ECG in GC as it might provide a potential therapeutic target for this subgroup of more aggressive tumors in the future.

Risk factors for positive sentinel lymph node, lymphatic or hematogenous dissemination over time in patients with cutaneous melanoma

The aim of the present study was to assess the influence of localization, age or sex and histopathological characteristics upon the chance of developing lymphatic or hematogenous metastatic spread over time, or a positive sentinel lymph node in cutaneous melanoma patients. Patients from the Department of Dermatology, County Emergency Hospital Cluj-Napoca (Cluj-Napoca, Romania), presenting with cutaneous melanoma confirmed histopathologically and a SPECT/CT or lymphoscintigraphic examination to detect the sentinel lymph node, were included in the present study. Our results revealed that Breslow index >2 mm [odds ratio (OR)=4.22, 95% confidence interval (CI) (1.12; 15.93)], presence of ulceration [OR=6.01, 95% CI (1.87; 19.35)], and positive sentinel lymph node [for at least one sentinel lymph node OR=3.58, 95% CI (1.06; 12.04)] were risk factors for hematogenous metastases. All these, except for the Breslow index >2 mm, were demonstrated to be a risk factor for lymphatic spread metastases over time. Ulceration and male sex also represented risk factors for a positive sentinel lymph node, men having a higher risk of developing sentinel lymph nodes than women [adjusted OR=2.27, 95% CI (1.00; 5.13)]. In conclusion, the predictors that influence the occurrence of lymphatic or hematogenous metastases may differ, ulceration and positive sentinel lymph node being common for both types of metastatic spread, while Breslow index being a significant predictor only for hematogenous metastases. Male sex and the presence of ulceration were demonstrated to be significant risk factors for positive sentinel lymph nodes.

Detection of Tumor Cell-Induced Platelet Aggregation and Granule Secretion.

Hematogenous metastatic spread of cancer is strongly dependent on and triggered by an intensive interplay of tumor cells with platelets. Immediately after entering the blood vascular system, tumor cells are surrounded by a platelet cloak, which protects them physically from shear stress and from attacks by the immune surveillance. Furthermore, tumor cell binding activates platelets, which in turn release growth factors and chemokines to recruit myeloid cells into the platelet/tumor cell microemboli, eventually create a permissive microenvironment in the early metastatic niche. Although the molecular mechanisms of tumor cells to activate platelets appear versatile being a matter of further research, interference with platelet activation turns out to be an attractive target to efficiently inhibit tumor metastasis. Some experimental assays are generally recognized to follow tumor cell-induced platelet activation (TCIPA), which provide an insight into the molecular mechanisms of TCIPA and allow searching for potential inhibitors. In this chapter, we describe the two most prominent experimental assays to follow TCIPA, namely platelet aggregation and platelet granule secretion, experimentally realized by dense granules´ ATP quantification. Although light transmission aggregometry and ATP detection from dense granule secretion are two age-old techniques, they are still highly relevant to provide reliable information concerning platelet activation status since all tumor cell-derived molecular triggers are covered and monitored in the experimental outcome.

Сomparative analysis of acridineacetate-containing compounds’ radio-sensitizing effect during malignant tumor experimental radiotherapy in a metastatic colorectal cancer model in balb/c mice

Radio-sensitizing compounds are used to increase radiotherapy effectivity and reduce negative pathologic systemic effects of radiation. These radio-sensitizers selectively pile-up in tumor tissue and enhance necrobiotic processes caused by irradiation. Unfortunately, broad introduction of radio-sensitizers into clinical practice is hampered by their low effectivity and substantial negative side effects. This what determines radio-sensitizers effectivity studies high actuality in a colorectal cancer mice model. The study involved 156 male BALB/c mice who had been transplanted with colorectal cancer. After the transplantation they were injected with acridineacetate containing compounds Cyclopherone and Procitol. A comparative assessment was undertaken of tumor growth dynamics and metastatic rate. A combination of gamma-irradiation with radio-sensitizing compounds produces a more substantial therapeutic effect according to all parameters of the experimental colorectal cancer growth and development than the isolated effect of radiotherapy or antitumor drugs. A considerable slowing down of tumor growth on the 14th and 28th day of the study was revealed as well as a reliable inhibition of lymphogenous and hematogenous metastatic spread of neoplasma during the entire study and a statistically valid increase of lifespan of the experimental animals. The radiosensitizing effect of Procitol in combination with radiotherapy turned out to be more substantial than that of Cyclopherone.

Heparanase in Cancer Metastasis- Heparin as a Potential Inhibitor of Cell Adhesion Molecules.

Tumor progression associated with hematogenous metastatic spread is a multistep process based on a cross-talk between tumor and stromal cells in a tumor microenvironment. In the blood circulation, tumor cells interact with blood cells through receptors such as selectin and integrins that promote tumor cells survival. At the metastatic sites, heparanase secreted by tumor or stromal cells is an important modifier of the tumor microenvironment while promoting tumor invasiveness and angiogenesis. Heparin, particularly low molecular weight heparin, is used for treatment of cancer patients with evidence of hypercoagulability. However, in preclinical studies heparins was shown to contain other biological activities that affect cancer progression including inhibition of heparanase, selectins and integrins. While ongoing clinical trials are assessing inhibition of heparanase on cancer progression, the remaining biological activities of heparins inhibiting cells adhesion, through selectins and integrins remains largely unexplored. This chapter addresses the potential role of heparins in oncology with respect to their anti-heparanase and anti-adhesive activities and aims to discuss aspects relevant for broader therapeutic application of heparins.

The diagnosis of metastatic uveal melanoma. What has changed during 10 years?

Uveal melanoma is the most common intraocular malignancy. Despite significant progress in the treatment of the primary tumor, uveal melanoma continues to be the most aggressive tumor due to its high potential for hematogenous metastatic spread. Uveal melanoma metastases are located mostly in the liver. Overall survival of patients with liver metastases is 4 to 15 months. There is no adjuvant systemic therapy to prevent metastases of uveal melanoma. In this paper, we aim to analyze achievement in diagnosis of metastatic uveal melanoma made during the last 10 years and to show advantages and disadvantages of the main diagnostic methods. The last years showed the emergence of understanding the differences between skin and uveal melanoma based on different molecular genetic abnormalities and different signaling pathways of these tumors. The success of target treatments for skin melanoma is not applicable to uveal melanoma. Skin and uveal melanoma responds differently to immune checkpoint inhibitors. Insufficient efficacy of systemic chemotherapy, immune and target therapy has made it necessary to develop techniques of local chemoembolization and radioembolization of uveal melanoma metastases. At present, there are no available assessment algorithms for the management of patients with uveal melanoma and suspected metastases. Ultrasonography, if seen as available techniques for radiology diagnostics, should be enhanced either by the intravenous contrast, or by qualifying computed tomography. The use of positron emission tomography / computed tomography has not demonstrated the maximal diagnostic value in identification of liver metastases, and, taken into account its high costs, low availability, and large radiation dose, it cannot be considered as a feasible method for every 6 months' application. The most effective diagnostic method is magnetic resonance imaging with the liver-specific contrast agents, due to the possibility to detect metastases of less than 1 cm in diameter. However, this technique has not been widely accepted clinically due to high costs, contraindications and a big proportion of false positive results. Current armamentarium of radiation techniques that could be potentially applicable for visualization of hepatic focal abnormalities is rather impressive and is being continuously updated; however, the problem of the metastatic uveal melanoma diagnosis remains unsolved.

Rapid evolution and biogeographic spread in a colorectal cancer

How and when tumoral clones start spreading to surrounding and distant tissues is currently unclear. Here we leveraged a model-based evolutionary framework to investigate the demographic and biogeographic history of a colorectal cancer. Our analyses strongly support an early monoclonal metastatic colonization, followed by a rapid population expansion at both primary and secondary sites. Moreover, we infer a hematogenous metastatic spread under positive selection, plus the return of some tumoral cells from the liver back to the colon lymph nodes. This study illustrates how sophisticated techniques typical of organismal evolution can provide a detailed, quantitative picture of the complex tumoral dynamics over time and space.

Molecular Mechanisms Regulating Organ-Specific Metastases in Epithelial Ovarian Carcinoma

Epithelial ovarian carcinoma is the most predominant type of ovarian carcinoma, the deadliest gynecologic malignancy. It is typically diagnosed late when the cancer has already metastasized. Transcoelomic metastasis is the most predominant mechanism of dissemination from epithelial ovarian carcinoma, although both hematogenously and lymphogenously spread metastases also occur. In this review, we describe molecular mechanisms known to regulate organ-specific metastasis from epithelial ovarian carcinoma. We begin by discussing the sites colonized by metastatic ovarian carcinoma and rank them in the order of prevalence. Next, we review the mechanisms regulating the transcoelomic metastasis. Within this chapter, we specifically focus on the mechanisms that were demonstrated to regulate peritoneal adhesion—one of the first steps in the transcoelomic metastatic cascade. Furthermore, we describe mechanisms of the transcoelomic metastasis known to regulate colonization of specific sites within the peritoneal cavity, including the omentum. Mechanisms underlying hematogenous and lymphogenous metastatic spread are less comprehensively studied in ovarian cancer, and we summarize mechanisms that were identified to date. Lastly, we discuss the outcomes of the clinical trials that attempted to target some of the mechanisms described in this review.

CT measurement of breast glandular tissue and its association with testicular cancer.

To evaluate the associations between breast glandular tissues diameters as determined by CT and b-hCG levels, histological types, tumour spread and prognosis in patients with testicular germ cell tumour. Ninety-four patients with pre-treatment CT scan and markers (b-hCG, AFP, LDH) were retrospectively collected. A radiologist measured diameters in all CT examinations and correlation between diameters and log (b-hCG) was assessed (Pearson's coefficient). The ability of measured diameters to predict lymphatic and distant haematogenous metastatic spread was evaluated (ROC curves). The associations between measured diameter cut-off values of 20 and 25mm and International Germ Cell Cancer Collaborative Group (IGCCCG) classification, lymphatic and distant haematogenous metastatic spread and histological subtypes were evaluated (chi squared test). Breast glandular diameters correlated to log(b-hCG) (r = 0.579) and predicted distant haematogenous metastatic spread (AUC = 0.78). Worse prognosis (intermediate or poor IGCCCG) was shown for 20mm (27.3 vs. 4.2%, p = 0.005) and 25mm (33.3 vs. 6.1%, p = 0.014). A diameter of 25mm was associated with non-seminoma (91.7 vs. 48.8%, p = 0.005). Breast glandular tissue diameters correlated with log(b-hCG) and predicted distant haematogenous metastases. Twenty and 25mm were associated with worse prognosis and 25mm was able to distinguish between seminoma and non-seminoma. • CT breast glandular tissue diameter correlates with log(b-HCG) • Gynaecomastia in CT is associated with worse prognosis • Gynaecomastia in CT is associated with non-seminoma histological subtype.

Does active smoking induce hematogenous metastatic spread in colon cancer?

BackgroundNo consensus exists on the influence of active smoking on the baseline staging of colon cancer patients. MethodsA cohort of colon cancer patients treated surgically at Massachusetts General Hospital (2004 to 2011) was reviewed. ResultsOf 1,071 patients, 563 reported ever smoking, among which 128 (12%) patients were current smokers. Ex-smokers and never smokers had similar rates of nodal (relative risk [RR] .9, P = .19) and metastatic disease (RR .96, P = .72), leading to comparable colon cancer–related mortality (RR 1.01, P = .95). Current smokers had similar rates of lymph node disease (RR 1.01, P = .88), but had significantly higher stage-adjusted odds of metastatic disease at presentation (odds ratio 2.57, 95% confidence interval 1.36 to 4.98, P = .005), in addition to higher stage-adjusted all-cause mortality (hazard ratio 1.44, P = .017). ConclusionsActive smoking was a stage-independent risk factor for baseline hematogenous metastasis and mortality. As this link was not present in former smokers, a potential healthcare benefit may be achieved in terms of baseline colon cancer presentation and outcomes through smoking cessation.

Effective and selective cell retention and recovery from whole blood by electroactive thin films.

Hematogenous metastatic spread causes most cancer patient deaths. Because circulating tumor cells (CTCs) are highly relevant to early metastatic spread, the capture or detection of these cells provides a diagnostic tool for patients with metastatic conditions. Herein, we demonstrate a programmable electroactive multilayered material platform with a smart electrically induced "switch" that captures CTCs from biological plasma with high efficiency and releases the captured cells flexibly. The released cells are still viable and proliferative, which facilitates the detection of trace levels of CTCs by amplification. Furthermore, the inherent rough characteristics of the nanoparticle-composed interface can promote capture efficiency and cell purification by integration with a simple microfluidic device. This elegant, inexpensive, and versatile platform for cell sorting and enrichment makes subsequent molecular and cell biological analysis achievable. The strategy has broad implications for favoring fundamental cancer biology research, for the diagnosis and monitoring of cancer individually, and for advanced intervention based on blood purification.

Circulating tumor cells: a multifunctional biomarker.

One of the most promising developments in translational cancer medicine has been the emergence of circulating tumor cells (CTC) as a minimally invasive multifunctional biomarker. CTCs in peripheral blood originate from solid tumors and are involved in the process of hematogenous metastatic spread to distant sites for the establishment of secondary foci of disease. The emergence of modern CTC technologies has enabled serial assessments to be undertaken at multiple time points along a patient's cancer journey for pharmacodynamic (PD), prognostic, predictive, and intermediate endpoint biomarker studies. Despite the promise of CTCs as multifunctional biomarkers, there are still numerous challenges that hinder their incorporation into standard clinical practice. This review discusses the key technical aspects of CTC technologies, including the importance of assay validation and clinical qualification, and compares existing and novel CTC enrichment platforms. This article discusses the utility of CTCs as a multifunctional biomarker and focuses on the potential of CTCs as PD endpoints either directly via the molecular characterization of specific markers or indirectly through CTC enumeration. We propose strategies for incorporating CTCs as PD biomarkers in translational clinical trials, such as the Pharmacological Audit Trail. We also discuss issues relating to intrapatient heterogeneity and the challenges associated with isolating CTCs undergoing epithelial-mesenchymal transition, as well as apoptotic and small CTCs. Finally, we envision the future promise of CTCs for the selection and monitoring of antitumor precision therapies, including applications in single CTC phenotypic and genomic profiling and CTC-derived xenografts, and discuss the promises and limitations of such approaches. See ALL articles in this CCR focus section, "Progress in pharmacodynamic endpoints."

A Donor Cornea With Metastatic Cells From a Cutaneous Malignant Melanoma

To describe the case of a donor cornea that showed hematogenous metastatic spread of cutaneous melanoma to the sclerocorneal limbus. Corneal tissue obtained from a donor with cutaneous malignant melanoma was evaluated for endothelial cell density, corneal transparency, and epithelial morphology. Subsequently, hematoxylin and eosin staining and immunohistochemical characterization using S100, HMB45, Melan-A, and CD34 antibodies were performed on the corneal sections. The corneal tissue was transparent with high endothelial cell density; it was graded as being suitable for transplantation according to the current eye bank criteria. However, the aggressiveness of the donor's cancer and the diffuse melanosis of the sclera led to the suspicion of malignant melanoma metastasis to the cornea. Histochemical analysis of the corneoscleral rim showed small aggregates rich in pigmented cells that were localized in cleft-like structures in the sclera, at the sclerocorneal interface and in the peripheral avascular portion of the cornea. The aggregates were positive for the melanocytic tumor markers S100, HMB45, and Melan-A; the rims of the clefts expressed the panvascular CD34 antigen, which was suggestive of neovascularization. Corneal tissue from a donor with malignant cutaneous melanoma displayed neoplastic lesions of melanocytic origin that had spread from a primitive melanoma through hematogenous routes to the sclerocorneal limbus. On the basis of this finding, we believe that having a metastatic cutaneous malignant melanoma could in some cases be reviewed as an exclusionary criterion for undergoing cornea transplantation.

Human chorionic gonadotropin (hCG) inhibits onset of preeclampsia in a “humanized” mouse model and restores uterine angiogenesis and regulatory T cells

During inflammation, E- and P-selectins appear on activated endothelial cells to interact with leukocytes through sialyl-Lewis x and sialyl-Lewis a antigens (sLex/a). These selectins can also interact with tumor cells in a sialyl-Lewis-dependent manner and for this reason, they are thought to play a key role in metastasis. Diverting the biosynthesis of sialyl-Lewis antigens toward nonadhesive structures is an attractive gene therapy for preventing the hematogenous metastatic spread of cancers. We have previously shown that transfection of α(1,2)-fucosyltransferase-I (FUT1) in Chinese hamster ovary (CHO) cells had a slight effect on the overall sialylation while the synthesis of sLEx was dramatically prevented. We herein delivered the gene of FUT1 by a human immunodeficiency virus-derived lentiviral vector to three human cancer cell lines including pancreatic (BxPC3), hepatic (HepG2), and colonic (HT-29) cancer cells. We found that on FUT1 transduction, all cells exhibited a dramatic decrease in sLex synthesis with a concomitant increase in Ley and Leb expression, without any detectable effect on the level of cell surface sLea antigens. In parallel, FUT1-transduced HT-29 and HepG2 cells, but not BxPC3 cells, failed to interact with E-selectin as assessed by E-selectin-binding assay or dynamic adhesion to activated endothelial cells. We show also that transduced FUT1 efficiently fucosylates the P-selectin ligand PSGL-1 without altering P-selectin binding. These results have important implications for understanding cell-specific reactions underlying the synthesis of selectin ligands in cancer cells and may provide a basis for the development of anti-metastatic gene therapy.

BEAM Trial: Lighting the Way Ahead?

BEAM Trial: Lighting the Way Ahead?

Abstract C44: Metastatic potential of prostate tumor cells is affected by hyaluronan metabolism.

Abstract Introduction: Accumulation of hyaluronan (HA) and elevation of the HA hydrolytic enzyme Hyal1 have been shown to promote prostate cancer progression. Immunohistochemical detection of HA and Hyal1 is diagnostic in biopsies and resected tissue, and predicts biochemical recurrence. We previously found that stable co-overexpression of Hyal1 and HAS, the HA biosynthetic enzyme, in 22Rv1 prostate tumor cells accelerates orthotopic tumor growth and spontaneous lymph node (LN) metastasis. Furthermore, the presence of cell-associated HA is required for adhesion of prostate tumor cells to bone marrow derived endothelial cells in vitro, suggesting surface HA could impact metastatic preference and/or efficiency of tumor cells by promoting arrest in specific vasculatures such as that of the bone. Here, we tested this hypothesis using intracardiac (IC) injections to model the hematogenous metastatic spread of tumor cells. Experimental Procedures: Male nu/nu, nude/SCID and NOD/SCID mice were injected via the left ventricle with 1×105 tumor cells in 0.1 ml HBSS containing IRDye 800CW carboxylate, a non-reactive near infrared (NIR) imaging agent, as an indicator of successful injection. Mice (n=75) were monitored over a 10 week period by non-invasive fluorescent optical imaging of dual probes in the 700 and 800 channels of the Pearl Impulse Small Animal Imager. Probes were designed to identify tumor tissue (IRDye 800CW conjugated to EGF, 2DG, or RGD), lymphatic tissue (IRDye 680RD hyaluronan (HA)) or bone tissue (IRDye 680RD BoneTag). Animals were imaged at the study endpoint and metastases noted. Tissues were excised, imaged ex vivo, and metastasis confirmed by histology. Results: PC3M-LN4 highly metastatic HA-encapsulated prostate tumor cells produced macrometastases at a higher rate (33%) after 10 wks than 22Rv1 HA-deficient cells (14%) in nu/nu and NOD/SCID mice. Suspect lesions were present in lung, bone, liver, adrenals, kidney, brain, and LN. Hyal1 overexpressing 22Rv1 cells were more frequently metastatic to brain and lungs and more rapidly growing, since that group exhibited a higher mortality rate. However, the frequency of metastasis observed in LN and bone was significantly higher for HAS transfectants, which also showed greater overall tumor burden in LN. Conclusions: Although Hyal1 expression promotes LN growth via lymphatic invasion from prostate, HAS expression and surface HA retention provide a greater LN colonization advantage for tumor cells already in circulation. Use of IRDye 800CW and IRDye 680RD optical imaging agents in a multiplex fashion provided strong evidence of macro- and micro-metastases along with additional detail relevant to the microenvironment upon intraoperative evaluation. In several instances the dual optical agent evaluation alerted us to the presence of suspect lesions subsequently confirmed by H&E staining. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C44.