RESUMO
Understanding the host response to oncolytic viruses is important to maximize their antitumor efficacy. Despite robust cytotoxicity and high virus production of an oncolytic herpes simplex virus (oHSV) in cultured human sarcoma cells, intratumoral (ITu) virus injection resulted in only mild antitumor effects in some xenograft models, prompting us to characterize the host inflammatory response. Virotherapy induced an acute neutrophilic infiltrate, a relative decrease of ITu macrophages, and a myeloid cell-dependent upregulation of host-derived vascular endothelial growth factor (VEGF). Anti-VEGF antibodies, bevacizumab and r84, the latter of which binds VEGF and selectively inhibits binding to VEGF receptor-2 (VEGFR2) but not VEGFR1, enhanced the antitumor effects of virotherapy, in part due to decreased angiogenesis but not increased virus production. Neither antibody affected neutrophilic infiltration but both partially mitigated virus-induced depletion of macrophages. Enhancement of virotherapy-mediated antitumor effects by anti-VEGF antibodies could largely be recapitulated by systemic depletion of CD11b(+) cells. These data suggest the combined effect of oHSV virotherapy and anti-VEGF antibodies is in part due to modulation of a host inflammatory reaction to virus. Our data provide strong preclinical support for combined oHSV and anti-VEGF antibody therapy and suggest that understanding and counteracting the innate host response may help enable the full antitumor potential of oncolytic virotherapy.
Assuntos
Vetores Genéticos/imunologia , Células Mieloides/imunologia , Neoplasias/imunologia , Vírus Oncolíticos/imunologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Animais , Anticorpos Monoclonais Humanizados/administração & dosagem , Anticorpos Monoclonais Humanizados/farmacologia , Bevacizumab , Antígeno CD11b/metabolismo , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Modelos Animais de Doenças , Feminino , Vetores Genéticos/administração & dosagem , Humanos , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Células Mieloides/metabolismo , Neoplasias/metabolismo , Neoplasias/terapia , Neovascularização Patológica/terapia , Terapia Viral Oncolítica , Sarcoma/imunologia , Sarcoma/metabolismo , Sarcoma/terapia , Simplexvirus/imunologia , Células Estromais/metabolismo , Células Estromais/virologia , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fator A de Crescimento do Endotélio Vascular/imunologia , Replicação Viral/efeitos dos fármacos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
PURPOSE: This study examined the influence of body fatness on body core temperature and heat loss responses during moderate-intensity exercise. METHODS: Nine men with lower body fat and eight men with higher body fat, matched for aerobic fitness, completed 1 h of recumbent cycling at the same absolute intensity in a warm environment (30 degrees C, 40% RH). Percent body fat was measured by hydrostatic weighing, using oxygen dilution to determine residual volume. Esophageal temperature (T(es)), mean skin temperature (T(sk)), and local sweat rate (m(sw)) were measured at rest and continuously during exercise while forearm blood flow (FBF) was measured at rest and every 10 min during exercise. RESULTS: The lower body fat and higher body fat groups were successfully matched for aerobic fitness, removing the influence of body fatness, given that V/O2(peak) was 50.72 +/- 7.34 and 50.43 +/- 5.01 ml x kg LBM(-1) x min(-1), respectively. When compared to lower body fat individuals, % body fat, body surface area (A(D)), and body mass were higher and A(D)/ mass was lower in higher body fat individuals. T(es), T(sk), FBF, m(sw), and the slope of m(sw):T(es) were not different between groups. Metabolic heat production was similar between the lower body fat (299.7 +/- 40.5 W x m(-2)) and higher body fat (288.1 +/- 30.6 W x m(-2)) subjects, respectively. Dry and evaporative heat loss, as well as heat storage during exercise, were not different between groups. CONCLUSION: These data suggest that there is no effect of body fatness on body core temperature or heat loss responses during moderate-intensity exercise in a warm environment.
Assuntos
Adiposidade/fisiologia , Regulação da Temperatura Corporal/fisiologia , Temperatura Corporal/fisiologia , Exercício Físico/fisiologia , Adulto , Feminino , Transtornos de Estresse por Calor/fisiopatologia , Humanos , Masculino , Consumo de Oxigênio/fisiologia , Adulto JovemRESUMO
BACKGROUND: Although disease remission can frequently be achieved for patients with neuroblastoma, relapse is common. The cancer stem cell theory suggests that rare tumorigenic cells, resistant to conventional therapy, are responsible for relapse. If true for neuroblastoma, improved cure rates may only be achieved via identification and therapeutic targeting of the neuroblastoma tumor initiating cell. Based on cues from normal stem cells, evidence for tumor populating progenitor cells has been found in a variety of cancers. METHODOLOGY/PRINCIPAL FINDINGS: Four of eight human neuroblastoma cell lines formed tumorspheres in neural stem cell media, and all contained some cells that expressed neurogenic stem cell markers including CD133, ABCG2, and nestin. Three lines tested could be induced into multi-lineage differentiation. LA-N-5 spheres were further studied and showed a verapamil-sensitive side population, relative resistance to doxorubicin, and CD133+ cells showed increased sphere formation and tumorigenicity. Oncolytic viruses, engineered to be clinically safe by genetic mutation, are emerging as next generation anticancer therapeutics. Because oncolytic viruses circumvent typical drug-resistance mechanisms, they may represent an effective therapy for chemotherapy-resistant tumor initiating cells. A Nestin-targeted oncolytic herpes simplex virus efficiently replicated within and killed neuroblastoma tumor initiating cells preventing their ability to form tumors in athymic nude mice. CONCLUSIONS/SIGNIFICANCE: These results suggest that human neuroblastoma contains tumor initiating cells that may be effectively targeted by an oncolytic virus.
Assuntos
Regulação Neoplásica da Expressão Gênica , Células-Tronco Neoplásicas/citologia , Neuroblastoma/metabolismo , Vírus Oncolíticos/metabolismo , Antígeno AC133 , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/biossíntese , Animais , Antígenos CD/biossíntese , Linhagem Celular Tumoral , Linhagem da Célula , Chlorocebus aethiops , Glicoproteínas/biossíntese , Humanos , Proteínas de Filamentos Intermediários/biossíntese , Proteínas de Neoplasias/biossíntese , Proteínas do Tecido Nervoso/biossíntese , Nestina , Peptídeos , Células-Tronco/metabolismo , Transcrição Gênica , Células VeroRESUMO
Malignant solid tumors remain a significant clinical challenge, necessitating innovative therapeutic approaches. Oncolytic viral therapy is a nonmutagenic, biological anticancer therapeutic shown to be effective against human cancer in early studies. Because matrix metalloproteinases (MMP) play important roles in the pathogenesis and progression of cancer, we sought to determine if "arming" an oncolytic herpes simplex virus (oHSV) with an MMP-antagonizing transgene would increase virus-mediated antitumor efficacy. We generated oHSVs that express human tissue inhibitor of metalloproteinases 3 (TIMP3) or firefly luciferase and designated them rQT3 and rQLuc, respectively. We evaluated the antitumor efficacy of these viruses against neuroblastoma and malignant peripheral nerve sheath tumor (MPNST) xenografts. Relative to rQLuc, rQT3-infected primary human MPNST and neuroblastoma cells exhibited equivalent virus replication but increased cytotoxicity and reduced MMP activity. In vivo, rQT3-treated tumors showed delayed tumor growth, increased peak levels of infectious virus, immature collagen extracellular matrix, and reduced tumor vascular density. Remarkably, rQT3 treatment reduced circulating endothelial progenitors, suggesting virus-mediated antivasculogenesis. We conclude that rQT3 enhanced antitumor efficacy through multiple mechanisms, including direct cytotoxicity, elevated virus titer, and reduced tumor neovascularization. These findings support the further development of combined TIMP-3 and oncolytic virotherapy for cancer.