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BACKGROUND Few studies have compared the features of thyroid cancer among races and ethnicities. We hypothesized that race and ethnicity may influence the frequency and features of thyroid malignancy in thyroid nodules. METHOD This was a retrospective chart review of patients between 2013 and 2020 who underwent thyroidectomy. RESULTS In the analysis of 2737 patients, thyroid cancer was less prevalent among Blacks (24.0% vs Whites 52.1%, Hispanics 58.7%, Asians 71.7%, and Others 57.9%, p

Quality of life (QoL) of endocrine surgery patients is an important patient outcome but the role of social determinants of health (SDH) on preoperative QoL is understudied.This study used preoperative data of 233 endocrine surgery patients participating in a longitudinal QoL study to examine the influence of SDH (patient-level and environmental) on preoperative QoL. Patient-level SDH was assessed with structured survey questions and environmental SDH with the Social Vulnerability Index. Multiple domains of QoL were assessed with the Patient-Reported Outcomes Measurement Information System-29 (PROMIS-29).The average age of the sample was 52.9 y and 76.8% were female, 10% were Hispanic, 55.8% were White, 32.6% were Black, 6.9% were Other, and 4.7% were Asian. Patients with patient-level SDH were more likely to have worse preoperative QoL in multiple PROMIS domains. Patients who lived in the most socially vulnerable areas had the same or better QoL scores in the PROMIS-29 domains than those living in less vulnerable areas. Minority race patients were more likely to have patient-level SDH and to live in the most vulnerable areas.This study is the first to our knowledge to examine the role of patient-level and environmental SDH on preoperative QoL among endocrine surgery patients. The results identified specific patient-level factors that could be used as the basis for interventions aimed to improve patients' QoL. Future studies that evaluate the role of preoperative SDH on long-term QoL and clinical outcomes would further enhance our understanding of the impact of SDH on patient wellbeing.

Background Thyroid surgery is becoming more common in the elderly as elderly population continues to grow. We aim to evaluate the relative risk of morbidity from thyroidectomy in patients greater than 75 y of age. Methods A retrospective analysis was performed for patients who were undergoing thyroidectomy between 2001 and 2018 in a multihospital network. A matched control group was selected with use of a propensity score, which was based on gender, ethnicity, type of surgery, insurance status, and comorbidities. The Charlson Comorbidity Index was used to quantify comorbidities. Total complications included both thyroid-specific and systemic complications. Results We identified 313 patients over the age of 75 y with a propensity score matched group of 313 patients. There was no difference between the percent female (73% versus 73%, P = 0.92), race composition (P = 0.91), insurance status (P = 0.99), percent undergoing total thyroidectomy (84% versus 84%, P = 0.91), and Charlson Index (2.6 versus 2.69, P = 0.70) of the two groups. Overall complications (4.8% versus 1.9%, P = 0.05) and thrombotic events (1.2 versus 0%, P = 0.04) were significantly higher but there was no statistically significant difference between postoperative emergency room visits (7% versus 6%, P = 0.61), readmissions (11.5% versus 8.6%, P = 0.18), cardiovascular (1.3 versus 0.6%, P = 0.61), pulmonary (3.2 versus 0.9%, P = 0.07), or neurologic complications (1.0 versus 0.3%, P = 0.34). No reoperations were noted in either group. Elder patients did have a longer length of stay (2.64 versus 1.29 d, P Conclusions Elderly patients did have a longer length of stay when compared to a matched younger population. Although there was a trend with higher complication rates in the elderly, those differences did not reach statistical significance.

Thymic hyperplasia is associated with Graves' disease, particularly in young patients. The degree of thymic transformation is minimal in most but not all patients. In the latter group radiological measurements of thyroid size and their change with treatment have rarely been reported. We present two patients with Graves' disease and relatively rapid resolution of thymic enlargement after successful treatment of their hyperthyroidism.Three patients with thyrotoxicosis secondary to Graves' disease and marked thymic enlargement were seen at our institution during a 2-year period. On computed tomography (CT) studies their volumes were 67, 81, and 54 cm(3). Thymic hyperplasia in the setting of Graves' disease was the diagnosis of exclusion. Two of the patients returned for follow-up after successful treatment of thyrotoxicosis as requested. On repeat CT their thymic volumes had decreased by 72% and 78%, respectively. Two types of histological modifications of the thymus have been described in association with Graves' disease, namely, thymic parenchyma hyperplasia and medullary lymphoid hyperplasia. The mechanisms underlying thymic transformation in patients with Graves' hyperthyroidism are not completely elucidated, but autoimmune processes underlying Graves' disease are presumed to play a role. The clinical course of our patients is consistent with earlier literature, indicating that thymic enlargement may occur in conjunction with Graves' hyperthyroidism, and that it usually resolves as hyperthyroidism is treated, but there is little quantitative pre- and posttreatment of hyperthyroidism data.Although every patient must be individually considered, it appears that thymic hyperplasia can be diagnosed in most Graves' hyperthyroid patients by considering the clinical context and appropriate radiologic studies such as CT. Raising awareness of the association of thymic hyperplasia in patients with Graves' hyperthyroidism and its resolution with the reversibility of the hyperthyroid state should prevent unnecessary thymic evaluation and surgery with its attendant risks.

Although most patients with differentiated thyroid cancer enjoy long survival, locoregional recurrence is common. In light of the high incidence of recurrence, how to best manage nodal recurrence is an important issue in the treatment of thyroid cancer patients. Surgery is one treatment option, although its effectiveness is hard to define because of the paucity of studies with large sample size and long follow-up. The percentage of patients rendered disease-free by surgery for nodal recurrence varies substantially from study to study as do the criteria that define freedom from disease. In the two most recent large studies of reoperation for nodal recurrence, remission was achieved in 23 % and 51 % of patients. Overall, surgery appears to be a reasonable option for managing nodal recurrence in the short term, but the optimal extent of surgery has not been determined. Long-term studies are needed to assess the impact of surgery on survival. Some advocate dissection of the compartment containing the recurrence only, while others advocate routine dissection of the central compartment for lateral nodal recurrence. Because many patients are left with persistent disease even after extensive surgery, a candid preoperative dialogue with the patient is essential to achieve realistic expectations of surgery.

BACKGROUND: Interleukin 1 (IL-1) is a pluripotent cytokine that promotes angiogenesis, tumor growth, and metastasis in experimental models; its presence in some human cancers is associated with aggressive tumor biology. The purpose of these studies was to characterize the role of IL-1 in human cancers and determine if inhibition of IL-1 via its receptor antagonist, IL-1Ra, alters tumor growth and metastatic potential. METHODS: IL-1 mRNA or protein levels were determined in clinical tumor samples, cancer cell lines, and xenografts using quantitative reverse transcription-PCR or ELISA. Biological activity of tumor-derived IL-1 protein was shown via induction of permeability across endothelial cell monolayers. The effects of recombinant IL-1Ra on tumor lines in culture (cell proliferation and IL-8 secretion) and in xenograft models (tumor growth, metastatic potential, and intratumoral levels of IL-8 and VEGF) were characterized. The effects of IL-1Ra-mediated regression of xenograft growth on angiogenic proteins (IL-8 and VEGF) were evaluated in an IL-1-producing melanoma (SMEL) xenograft model. RESULTS: IL-1 mRNA was highly expressed in more than half of all tested metastatic human tumor specimens including non-small-cell lung carcinoma, colorectal adenocarcinoma, and melanoma tumor samples. Constitutive IL-1 mRNA expression was identified in several cancer cell lines; tumor supernatant from these cell lines produced a significant increase in endothelial cell monolayer permeability, a hallmark event in early angiogenesis, in an IL-1-dependent manner. Moreover, systemic recombinant IL-1Ra resulted in significant inhibition of xenograft growth and neovessel density of IL-1-producing, but not non-IL-1-producing, tumor cell lines. Subsequent analysis of SMEL, a melanoma cell line with constitutive IL-1 production, showed that neither exogenous IL-1 nor IL-1Ra altered tumor cell proliferation rates in vitro. Gene expression analyses of IL-1Ra-treated SMEL xenografts showed a >3-fold down-regulation of 100 genes compared with control including a marked down-regulation of IL-8 and VEGF. CONCLUSIONS: These data show that the IL-1 gene is frequently expressed in metastases from patients with several types of human cancers. IL-1Ra inhibits xenograft growth in IL-1-producing tumors but has no direct antiproliferative effects in vitro; decreased tumor levels of IL-8 and VEGF may be an early surrogate of IL-1Ra-mediated antitumor activity. IL-1Ra may have a role alone or with other agents in the treatment of human cancers.

We are reporting the case of an ambulatory young woman with a 10-year history of recurrent venous thrombosis who presented to us with diffuse intravascular coagulation (DIC). After excluding the recognized causes of DIC, we examined the possibility that her clinically quiescent ulcerative colitis might be the underlying stimulus. We documented sepsis-range endotoxemia in this patient at a time when she was afebrile and had a normal C-reactive protein level. In vitro her serum upregulated tissue factor in cultured endothelial cells. We postulate that she had become tolerant to the systemic effects of endotoxin leaking from her inflamed colon but that the endotoxin stimulated her endothelium and/or monocytes to produce tissue factor that made her intensely hypercoagulable. Her prothrombotic state may have been compounded by the fact that she was heterozygous for prothrombin G20210A and that her plasma clotting time demonstrated resistance to activated protein C.

TNF is a cytokine with potent antitumor activity in murine models and when administered clinically via regional perfusion. There is substantial evidence that this antitumor activity depends in large part on TNF's procoagulant effect on tumor neovasculature, which is mediated by induction of endothelial cell tissue factor (TF), a component of the extrinsic clotting cascade. In regional perfusion of a cancer-bearing limb or organ, TNF is always administered under hyperthermic temperatures; however, little is known about the effect of hyperthermia on TNF-mediated procoagulant activity in endothelium. We examined the effects of hyperthermia on TNF-mediated procoagulant activity in human umbilical vein endothelial cells (HUVECs). HUVECs were exposed to TNF at normothermic (37 degrees C) and hyperthermic (41 degrees C) temperatures for 90 min, then assayed for clotting activity, TF protein production and mRNA production of TF and tissue factor pathway inhibitor-2 (TFPI-2), an endogenous inducible inhibitor of TF activity in HUVECs. TNF treatment at 41 degrees C significantly reduced clotting activity, TF protein and mRNA as well as TFPI-2 mRNA compared to treatment at 37 degrees C. These data show that hyperthermia significantly reduces the procoagulant effects of TNF on endothelial tissue compared to normothermia, which may have important clinical implications for the use of TNF in regional perfusion.

Spinal metastases (SMs) due to thyroid cancer (TC) are associated with significantly reduced quality of life. The goal of this study is to analyze the clinical manifestations, presentation, and treatments of TC SMs, and to describe specific features of SMs associated with different TC types.A retrospective analysis of 202 TC SM patients treated at Medstar Washington Hospital Center (37) and collected from the literature (165) was performed.The mean age of patients with SMs was 56.9±14.7 years, and the female-to-male ratio was 2.1:1. Of all patients, 29% (28% of follicular thyroid cancer [FTC] and 37% of papillary thyroid cancer [PTC]) had SMs only. Twenty-nine percent of all patients and 54% of patients with single-site SMs had neither bone non-SMs nor solid organ metastases at the time of presentation. Thirty-five percent of patients had SMs as an initial presentation of TC. TC patients presenting with SMs had a lower rate of other bone and visceral involvement compared with patients whose SMs were diagnosed at the time of thyroid surgery or during follow-up (p0.05). SMs were more often the initial manifestation of FTC (41% vs. 24%), while PTC SMs were more commonly diagnosed after TC diagnosis (76% vs. 59%; p0.05). PTC SMs were more frequently diagnosed as synchronous (63% vs. 36% in FTC) versus FTC SMs that developed as metachronous metastases (64% vs. 37% in PTC; p0.01). All FTC SMs developed within 82 (0-372) months and all PTC SMs within 35 (0-144) months (p0.01). In FTC SMs as TC manifestation, solid organ metastases involvement was less common than in FTC SMs that were found after TC diagnosis (34% vs. 67%; p0.01); multisite FTC SMs compared to solitary FTC SMs were associated with the development of other bone nonspinal metastases (82% vs. 30%; p0.01) and solitary organ metastases (65% vs. 41%; p0.01). These correlations were not observed in PTC SMs. FTC patients often had neural structure compression (myelopathy/radiculopathy; 72% vs. 36% in PTC), while PTC patients frequently were asymptomatic (38% vs. 5% in FTC; p0.01). FTC SMs more commonly were (131)I-avid (p0.01). FTC patients required surgery more frequently (72% vs. 55% in PTC; p0.05).Our study reveals that a significant part of TC SMs patients have solitary spinal involvement at the time of presentation and may be considered for aggressive treatment with the intention to improve quality of life and survival. FTC SMs and PTC SMs appear to have distinct presentations, behavior, and treatment modalities, and should be categorized separately for treatment and follow-up planning.

The spine is the most common site of bone metastases due to thyroid cancer, which develop in more than 3% of patients with well-differentiated thyroid cancer. Nearly half of patients with bone metastases from thyroid cancer develop vertebral metastases. Spinal metastases are associated with significantly reduced quality of life due to pain, neurological deficit, and increased mortality.Treatment options for patients with thyroid spinal metastases include radioiodine therapy, pharmacologic therapy, and surgical treatments, with recent advances in radiosurgery and minimally invasive spinal surgery as well. Therapeutic interventions require a multidisciplinary approach and aim to control pain, preserve or improve neurologic function, optimize local tumor control, and improve quality of life. We have proposed a three-tiered approach to the management and practical algorithms for patients with spinal metastases from thyroid carcinoma.The introduction of novel and improved techniques for the treatment of spinal metastases has created the opportunity to significantly improve control of metastatic tumor growth and the quality of life for the patients with spinal metastases from thyroid cancer. In order for these options to be effectively used, a multidisciplinary approach must be applied in the management of the patients with thyroid spinal metastases.

Viruses used for gene therapy are usually genetically modified to deliver therapeutic transgenes and prevent viral replication. In contrast, replication-competent viruses may be used for cancer therapy because replication of some viruses within cancer cells can result in their destruction (oncolysis). Viral ribonucleotide reductase expression is defective in the HSV1 mutant hrR3. Cellular ribonucleotide reductase, which is scarce in normal liver and abundant in liver metastases, can substitute for its viral counterpart to allow hrR3 replication in infected cells. Two or three log orders more of hrR3 virions are produced from infection of colon carcinoma cells than from infection of normal hepatocytes in viral replication assays. This viral replication is oncolytic. A single intravascular administration of hrR3 into immune-competent mice bearing diffuse liver metastases dramatically reduces tumor burden. hrR3-mediated tumor inhibition is equivalent in immune-competent and immune-incompetent mice, suggesting that viral oncolysis and not the host immune response is the primary mechanism of tumor destruction. HSV1-mediated oncolysis of diffuse liver metastases is effective in mice preimmunized against HSV1. These results indicate that replication-competent HSV1 mutants hold significant promise as cancer therapeutic agents. Yoon, S. S., Nakamura, H., Carroll, N. M., Bode, B. P., Chiocca, E. A., Tanabe, K. K. An oncolytic herpes simplex virus type 1 selectively destroys diffuse liver metastases from colon carcinoma.

Herpes simplex virus type 1 (HSV 1) vectors are under investigation for use in gene therapy for colorectal cancer liver metastases. Approximately 60% of colorectal cancers possess p53 mutations, and p53 mutations can cause tumor cell resistance to radiation therapy and chemotherapy. p53 is also known to colocalize with at least one HSV 1 protein and influence HSV 1 gene expression. The purpose of this study was to determine if the loss or mutation of p53 in tumor cells alters the cytotoxicity of HSV 1 vectors. HSV 1 vector-mediated in vitro cytotoxicity assays were performed using stable transfectants of SAOS-2-LM2 cells and WiDr cells that express no p53, wild-type p53, mutant p53, or both wild-type p53 and mutant p53. All stable transfectants were equally susceptible to HSV 1 vector cytotoxicity, and cell lines with mutant p53 were not resistant to HSV 1 vectors. These results provide additional rationale for the application of HSV 1 vector gene therapy for colorectal cancer.

The liver is a common site of metastases from solid tumors and the primary site for a number of malignancies. Cure rates for primary and metastatic liver tumors are low. Gene therapy is a novel approach to liver cancer that seeks to exploit differences between tumor cells and hepatocytes to achieve tumor eradication with preservation of normal hepatic parenchyma. Numerous strategies for tumor specific cytolysis are reviewed.

Entry of herpes simplex virus (HSV) into tumor cells results in viral gene expression followed by cellular lysis. Attenuated HSVs selectively destroy tumors with sparing of surrounding normal tissue. HSV encodes a thymidine kinase (TK) that converts ganciclovir to a toxic metabolite. This metabolite may be transferred between cells and lead to the death of neighboring uninfected cells, termed bystanders. We sought to determine if HSV-mediated oncolysis is enhanced by ganciclovir treatment. In addition, we examined bystander killing in cocultures of TK transfectants and parental cells. hrR3, an attenuated HSV, expresses TK. The 50% lethal dose of hrR3 for a rat gliosarcoma (9L) and three human colorectal carcinomas (HT29, KM12C6, and KM12L4) was determined. Cells were infected with a 50% lethal dose of hrR3, followed by treatment with ganciclovir, and then cell survival was quantitated. In separate experiments 9L and HT29 cells were transfected with TK. Parental cells and TK transfectants were cocultured in various ratios, in the presence of ganciclovir, and cell survival was quantitated. hrR3-mediated oncolysis was enhanced by ganciclovir in the gliosarcoma but not in the three colorectal carcinomas. Cocultures of both 9L and HT29 parental cells with their corresponding TK transfectants demonstrated bystander killing. The mortality of 9L cocultures was 54% greater than that predicted for exclusive killing of transfectants. HT29 mortality was 8% greater than predicted. The ability of ganciclovir to augment hrR3-mediated oncolysis varies significantly between tumor cells lines. The extent of ganciclovir-mediated killing of neighboring nontransduced parental cells similarly varies. Consideration should be given to these factors in the design of gene therapy strategies using HSV vectors as oncolytic agents.

OBJECTIVE: The authors determined whether a recombinant herpes simplex virus (HSV) vector could destroy human colon carcinoma cells in vitro and whether the vector would selectively replicate in colon carcinoma liver metastases but not surrounding hepatocytes in vivo. BACKGROUND: The HSV vector hrR3 is defective in the gene encoding ribonucleotide reductase and contains the lacZ reporter gene. Ribonucleotide reductase is expressed in actively dividing cells and generates deoxyribonucleotides for DNA synthesis. hrR3 replicates only in actively dividing cells that can provide ribonucleotide reductase in complementation, but not in quiescent cells such as normal hepatocytes. METHODS: hrR3-mediated lysis of HT29 human colon carcinoma cells was first determined in vitro. For in vivo studies, athymic BALB/c nude mice underwent intrasplenic injection of HT29 and intrasplenic injection of hrR3 7 days later, and were killed 7 days after viral injection. Their livers were examined histochemically for lacZ expression. RESULTS: All the HT29 cells were destroyed in vitro when hrR3 was added at a titer of 1 plaque-forming unit per 10 tumor cells. One hundred one of 105 tumor nodules examined in liver sections from mice treated by intrasplenic injection of hrR3 demonstrated lacZ expression. Minimal beta-galactosidase activity was present in normal liver. CONCLUSIONS: The hrR3 HSV vector effectively destroys HT29 human colon carcinoma cells at very low multiplicities of infection. Differential expression of ribonucleotide reductase between liver metastases and normal liver allows hrR3 to selectively replicate in tumor cells with minimal replication in surrounding normal liver. Further investigation of HSV-based vectors as oncolytic agents for liver metastases is warranted.

Combined therapy appears to be an effective method for the treatment of peritoneal mesothelioma. It can be performed safely and with acceptable morbidity. The WCI has reported a median overall survival of 67 months with a projected 3-year survival rate of 64% in 68 patients treated with various combinations of cytoreduction, CHPP, and intraperitoneal chemotherapy. The NCI has evaluated 49 patients who received combined therapy for peritoneal mesothelioma and found a median overall survival of 92 months with a projected overall 3-year survival of 59%. In both cases the outcomes are remarkably better than those in most previous reports of treatments for peritoneal mesothelioma, though these results need to be interpreted with caution, due to the short follow-up and selected study population.

IL-1β is a pleotropic cytokine that may mediate increased procoagulant activity and permeability in endothelial tissue during inflammatory conditions. The procoagulant effects of IL-1β are mediated through induction of tissue factor (TF) but its alterations on vascular permeability are not well characterized. We found that IL-1β induced a rapid and dose-dependent increase in TF activity in human umbilical vein endothelial cells (ECs) under routine culture conditions. However, IL-1β caused a rapid and marked increase in permeability across confluent EC monolayers using a two-compartmentin vitromodel only in the presence of factor VIII-deficient plasma that was completely abrogated by neutralizing anti-TF antibody pre-treatment.In vitropermeability was associated with loss of EC surface expression of VE-cadherin and contraction of F-actin cytoskeletal elements that resulted in EC intercellular gap formation. These data demonstrate that IL-1β induces marked changes in permeability across activated endothelium via a TF dependent mechanism and suggest that modulation of TF activity may represent a strategy to treat various acute and chronic inflammatory conditions mediated by this cytokine.

Interferon γ–inducible protein 10 (IP-10) has antitumor effects in various murine models. The IP-10 receptor has two distinct splice variants, CXCR3A and CXCR3B, that have paradoxical effects after ligand-receptor interaction. To characterize the putative antiangiogenic effects of IP-10, we measured proliferation rates and apoptosis in human umbilical vein endothelial cells (HUVECs), fibroblasts, and A375 melanoma or WIDR adenocarcinoma cell lines after exposure to the recombinant protein. CXCR3A (activating) and CXCR3B (inhibitory/proapoptotic) messenger RNA (mRNA) expression levels in fibroblasts, 2 human tumor cell lines, T lymphocytes, and HUVECs of varying cell densities were characterized. IP-10 resulted in dose-dependent and selective inhibition of proliferation and countered the proliferative effects of vascular endothelial growth factor in HUVECs but did not affect fibroblasts or 2 human tumor cell lines. In addition, IP-10 resulted in potent and selective induction of apoptosis in HUVECS but had no effect on fibroblasts or A375 melanoma. Confluent HUVECs had a predominance of mRNA for the CXCR3B splice variant by reverse transcriptase-polymerase chain reaction, and the ratio of CXCR3B to CXCR3A mRNA was >40 in HUVECs, compared with ≤10 in the other cell types. Moreover, CXCR3B mRNA levels were significantly higher in proliferating compared with confluent HUVECs. In vivo, systemic IP-10 administration resulted in slower A375 xenograft growth rates compared with control-treated animals, and immunohistochemical staining showed decreased microvessel density in xenografts of IP-10–treated mice. IP-10 has antiangiogenic properties and selective effects on endothelial tissue that may be secondary to higher levels of the CXCR3B inhibitory/proapoptotic receptor in that cell type, particularly in its actively proliferating state.

Interleukin (IL)-1 is a pleiotropic inflammatory cytokine that promotes angiogenesis and enhances tumor growth and metastases. We evaluated the effects of IL-1 receptor antagonist (IL-1ra) on tumor growth and metastases in human melanoma xenografts. We selected two human melanoma lines (SMEL and PMEL) with differential (high versus low, respectively) constitutive production of IL-1 by ELISA. The IL-1ra gene was isolated from monocyte RNA by PCR and retrovirally transduced into SMEL and PMEL. In vitro cell proliferation was evaluated using a WST-1 assay. Athymic nude mice received s.c. or i.v. injection with parental, vector-transduced, or IL-1ra-transduced melanoma cells, and tumor growth, lung metastases, and histology were characterized. IL-1 was produced by SMEL in vitro and ex vivo (117 and 67 pg/ml/10(6) cells/24 h, respectively), but not by PMEL (15 and 0 pg/ml/10(6) cells/24 h, respectively). Neither made IL-1ra natively. Gene-transduced cell lines secreted1000 pg/ml/10(6) cells/24 h of IL-1ra by ELISA. In vitro proliferation of each parental cell line was comparable to the proliferation rate of each transduced cell line. IL-1ra-transduced SMEL (SMEL/IL-1ra) showed significantly slower tumor growth compared with null-transduced and parental cell lines (P0.001, ANOVA-Bonferroni/Dunn). There was no difference in growth rates between PMEL and IL-1ra-transduced PMEL (PMEL/IL-1ra). A mixing study of SMEL and SMEL/IL-1ra showed significant inhibition of tumor growth at various ratios (P0.001, ANOVA-Bonferroni/Dunn). There were significantly fewer lung metastases with SMEL/IL-1ra versus SMEL (P0.002). IL-1ra decreases in vivo growth and metastatic potential of a human melanoma xenograft that constitutively secretes IL-1. This effect may be exploitable using clinically available IL-1ra for the treatment of human cancers.

Most cancer gene therapies using viruses have focused on replication-defective adenovirus, adeno-associated virus, retrovirus, vaccinia virus, and herpes simplex virus 1 (HSV-1). When using such replication-defective viruses, antineoplastic effects are achieved by delivery and expression of therapeutic transgenes. Although replication-defective viruses are used in a majority of current cancer gene therapy trials, 1 this approach has important drawbacks. Therapeutic transgene delivery is nonselective: both normal and cancer cells are infected. Replication-defective viruses are also unable to spread progeny virion to cells that were not initially infected. An alternative strategy exploits viral replication for tumor destruction, whereby infection of tumor cells by virus leads to cell destruction and simultaneous release of progeny virion that can infect adjacent tumor cells. 2 In this strategy, antineoplastic efficacy is dependent on viral replication; accordingly, it is important to maintain robust viral replication in neoplastic cells and simultaneously attenuate viral replication in nonneoplastic cells. Replication-conditional viruses can also deliver therapeutic transgenes to improve antineoplastic efficacy beyond that achieved by replication alone. 3 Tumor infection with replication-conditional viruses leads to longer transgene expression and better distribution throughout the tumor compared to tumor infection with replication-defective viruses. 4 Several viruses have been examined for their utility as replication-conditional oncolytic viruses, including HSV-1, 5 vaccinia virus, 6 Newcastle disease virus, 7 adenovirus, 8 and reovirus. 9 We have previously demonstrated preferential replication of an HSV-1 mutant hrR3 in tumors rather than normal tissue following intravascular administration. 10,11 hrR3 replication and oncolysis are attenuated in normal or quiescent cells because it is deficient in expression of viral ribonucleotide reductase (infected cell protein 6 [ICP6]). 12 The selectivity of hrR3 for liver tumors is related to significantly higher expression of ribonucleotide reductase and higher intracellular nucleotide pools in most tumors compared to surrounding normal tissues. 13 However, there are disadvantages to developing ICP6-defective HSV-1 mutants for clinical studies. ICP6-defective HSV-1 mutants are most effective against replicating cells, and accordingly quiescent cancer cells may be less susceptible to viral oncolysis. In addition, some virus may reach sites outside the liver despite regional administration, 14 and normal tissues with high replicative activity may be susceptible to infection and cytolysis by an ICP6-defective HSV-1 mutant. The normal tissues that are at greatest risk are those with high replicative activity, such as gastrointestinal mucosa and bone marrow. We have looked to manipulate viral genes that are critical for robust HSV-1 replication but, unlike viral ribonucleotide reductase, have no cellular homologues and whose function is poorly complemented by normal cells. The immediate-early ICP4 gene product is required for HSV-1 replication in cell culture. 15 Cells do not efficiently complement HSV-1 deficiency of ICP4, and accordingly, ICP4-defective HSV-1 mutants display markedly attenuated replication. The ICP4 gene is therefore a good candidate gene to regulate to achieve replication preferentially in cancer cells. Another good candidate gene to regulate is γ134.5, which is also required for robust HSV-1 replication. The γ134.5 gene product interacts with a cellular protein phosphatase to dephosphorylate elongation initiation factor 2α (eIF-2α) and permit cellular (and viral) protein translation to proceed. 16,17 Regulation of γ134.5 expression has been demonstrated by others to be an effective strategy to regulate HSV-1 lytic replication. 18 We examined a strategy in which ICP4 expression and γ134.5 expression are regulated by heterologous transcriptional regulatory elements for tumor-associated antigens. We first examined transcriptional regulatory elements for carcinoembryonic antigen (CEA) and MUC1/DF3 because they are overexpressed in a wide variety of epithelial cancers. We isolated HSV-1 mutants in which either ICP4 or γ134.5 expression is regulated by transcriptional regulatory elements for CEA and MUC1.

Thousands of patients die annually from unresectable metastatic or primary hepatic cancers confined to liver. Isolated hepatic perfusion (IHP) is a regional treatment strategy in which the vascular supply to the liver is isolated and perfused with a therapeutic regimen using an extracorporeal circuit consisting of a reservoir, heat exchanger, and oxygenator. Drug doses that would cause severe toxicities if delivered systemically can be confined to the liver by isolated hepatic perfusion, resulting in the ability to intensify treatment to the cancer-burdened region of the body. Agents and mechanisms commonly used in IHP include melphaIan, hyperthermia, and tumor necrosis factor. IHP appears to be efficacious for patients with advanced disease, as reflected by large tumor size, high number of lesions, or significant overall tumor burden in the liver. In addition, responses are observed for patients whose cancer is refractory to systemic and hepatic arterial infusion chemotherapy. Recent clinical trials have demonstrated that IHP has anti-tumor efficacy against primary hepatic neoplasms and metastases from various primary tumors, such as colorectal carcinoma, ocular melanoma, and neuroendocrine tumors. Current studies demonstrate that combining hepatic arterial infusion with floxuridine after IHP for patients with colorectal cancer metastases is associated with significant and durable response rates. Continued clinical evaluation is warranted for the use of IHP in the treatment of unresectable liver metastases.

OBJECTIVE: The authors investigate the efficacy of hrR3, a viral vector derived from herpes simplex virus type 1 (HSV 1), in destroying colon carcinoma cells in vitro and in vivo. The effect of adding the prodrug ganciclovir in combination with hrR3 infection also is assessed. SUMMARY BACKGROUND DATA: Most cancer gene therapy strategies use viral vectors that are incapable of replication. The HSV 1 vector hrR3 is capable of replication, and its replication is cytotoxic to cells. hrR3 also possesses the HSV-thymidine kinase gene, which converts ganciclovir into a toxic metabolite. Thus, the addition of ganciclovir to hrR3-infected cells may enhance the ability of hrR3 to destroy tumor cells. To increase specificity for tumor cells, hrR3 has a mutated ribonucleotide reductase gene and replicates selectively in cells with high levels of endogenous rbonucleotide reductase. Actively dividing cells such as tumor cells have high levels of endogenous ribonucleotide reductase for synthesis of DNA precursors. The authors are interested in the use of HSV 1 vectors to treat liver metastases from colorectal cancer. METHODS: Ribonucleotide reductase expression in several colon carcinoma cell lines and in primary cultures of human hepatocytes was determined by Western blot analysis. hrR3-mediated cytotoxicity in the colon carcinoma cell lines was determined using an in vitro assay. The human colon carcinoma cell line HT29 was injected into the flanks of nude mice followed by intratumoral injection of hrR3. Tumor growth rate was assessed with and without the addition of intraperitoneal ganciclovir. RESULTS: Ribonucleotide reductase levels in colon carcinoma cell lines are much higher than in primary cultures of human hepatocytes. hrR3 efficiently destroys colon carcinoma cell lines in vitro. A single intratumoral injection of hrR3 into HT29 flank tumors significantly reduces tumor growth rate, and the administration of ganciclovir has no additive effect. CONCLUSIONS: The inherent cytotoxicity of hrR3 replication effectively destroys colon carcinoma cells in vitro and in vivo. This cytotoxicity is not enhanced in vivo by the addition of ganciclovir. In the future, more efficacious and selective HSV 1 vectors may be useful in the treatment of cancer.

Background. Medullary thyroid cancer (MTC) is associated with RET mutations and activation of PI3K/AKT and MEK/ERK signaling pathways. Heat shock protein 90 (HSP90) contributes to functional stabilization of PI3K/AKT signaling and is required for growth factor inducible activation of the MEK/ERK pathway. A recent study showed that HSP90 is a molecular target for HIV protease inhibitor Nelfinavir (NFV). Treatment with NFV is associated with growth inhibition and induction of apoptosis in breast, ovarian and prostate cancer cell lines. Objectives. We hypothesized that NFV-inducible inhibition of HSP90 would lead to down-regulation of PI3K/AKT and MAPK/ERK signaling pathways and induction of apoptosis in MTC cells. Methods. MTC-derived cells (TT cells) harboring the RET C634W mutation were treated with NFV at concentrations from 1μM to 10μM. Activation of MAPK/ERK and AKT/ mTOR/p70S6K/pS6 signaling was assessed by Western blot using specific phospho-antibodies. Apoptosis was determined by Western Blot with antibody against cleaved caspase 3. Cell growth and viability were determined by Vi-Cell analyzer and Alamar blue assay. HSP90 expression was examined in 12 human MTC tumor samples by immunostaining. HSP90 expression was correlated with pAKT and pERK level. Results. Treatment with NFV (1μM to 10μM for 48 hours) did not affect expression of HSP90 or levels of total AKT and ERK. In contrast, pAKT and pERK were inhibited in TT cells after treatment with NFV (5μM and 10 μM). Since both AKT and ERK pathways contribute to regulation of p70S6K/pS6 activity, we examined activation of this signaling axis. NFV (10μM) decreased the level of p-p70S6K, p-pS6 and down-regulated Cyclin D1. Growth experiments demonstrated that the number of viable cells decreased by 30% and 85% after treatment with NFV at concentration 5μM and 10μM, respectively. Microscopy showed that exposure to NFV was associated with cell rounding and nuclear fragmentation, suggesting induction of apoptosis. Western blot analysis revealed that NFV induced caspase 3 cleavage in TT cells. In addition, treatment with NFV was associated with induction of γH2AX, indicating DNA damage. Immunostaining of human MTC showed increased expression of HSP 90 in 5/12 of examined MTCs compared to corresponding normal thyroid tissue. The highest level of cytoplasmic HSP90 was detected in 3 cases of MTC harboring RET mutation, and co-localization staining showed that cells with high level of HSP90 expression also exhibited prominent pAKT and pERK. Conclusion. Treatment of MTC cell with clinically achievable concentrations of NFV results in down-regulation of AKT and ERK signaling and inhibition of MTC cell growth. Expression of HSP90 in human tumors suggests that it has a role in maintenance of anti-apoptotic signaling in MTC cells. Together these data support the potential utility of NFV for the treatment of patients with MTC. Citation Format: Yevgeniya Kushchayeva, Aneeta Patel, John Costello, Vasyl Vasko, Nancy Carroll, Rubie Sue Jackson, Kenneth Burman, Lisa Boyle. The HIV protease inhibitor Nelfinavir inhibits medullary thyroid cancer cell growth. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2124. doi:10.1158/1538-7445.AM2013-2124

Thrombosis is a recognized complication of inflammatory bowel disease (IBD). In contrast disseminated intravascular coagulation (DIC) has rarely been described with IBD, and then only during periods of intense disease activity or during colectomy. However, we recently encountered a young woman with a 3-year history of ulcerative colitis (UC) and a 10-year history of recurrent thrombosis who presented with DIC when her disease appeared to be well controlled. For the previous two months she had been forced to discontinue her usual warfarin because of extreme over anticoagulation and extensive ecchymoses. Otherwise she felt well and was working full time. When she presented to us, the following laboratory data were obtained: pro time 18.7s, aPTT 43.6s, fibrinogen 56 mg/dL, D-dimer 8–16 μg/mL, platelet count 123K/μL, thrombin-antithrombin complexes 47 μg/mL (nl, 1.3–4.0), plasma endotoxin 6.3 EU/mL (control 0.1), positive prothrombin G20210A heterozygosity, negative FVLeiden, but positive APC resistance. Antithrombin, protein C, protein S, and homocysteine were within normal limits. Extensive imaging studies failed to reveal evidence of malignancy or venous malformations. The patient was treated with 1 mg/kg/d enoxaparin with resolution of her ecchymoses and normalization of her laboratory values. Subsequently we exposed cultured human umbilical vein endothelial cells (HUVECs) to our patient’s serum and assayed HUVEC lysates for tissue factor. After a 2 hour incubation a 20% dilution of her serum increased the tissue factor in the HUVECs to a level twice that observed with serum from a normal donor. Furthermore, the effect of her serum was negated by the addition of 1 μg/mL polymyxin-B, which specifically inhibits endotoxin. Discussion: Prothrombin G20210A, APC resistance, and UC are individually associated with an increase risk of thrombosis. However, the combination of all 3 factors in our patient apparently led to an intense degree of hypercoagulability that provoked not only recurrent thrombosis but DIC. Furthermore, the contribution of UC to her hypercoagulability may have been mediated by endotoxin diffusing from her inflamed bowel and upregulating tissue factor in her endothelium and/or monocytes. Because this patient’s recurrent thromboses predated her diagnosis of UC by 7 years, we suggest that occult IBD should be considered in the evaluation of idiopathic thrombophilia.