RESUMO
High-risk HPV E6 and E7 oncogenes are an ideal targeting gene for treatment of cervical cancer. In this paper, we introduce researches on cancer-immunotherapy targeting HPV E7 through mucosal immunity and E6/E7-targeting siRNA therapy using PEGylated polymeric micelles. Therapeutic HPV vaccine has also attracted attention as a cancer immunotherapy agent. We have found homing of Integrin ß7-positive intestinal mucosal lymphocyte on the cervical mucosa. In this study, we generated a novel therapeutic vaccine; an HPV E7-expressing Lactobacillus casei (LacE7) to induce anti-HPV cellular immunity directly to intestinal mucosa. Cervical lymphocytes (CxLs) and peripheral blood mononuclear cells (PBMCs) were counted E7 specific INFγ-producing cells (E7 cell-mediated immune responses: E7-CMI) by ELISPOT assay. We confirmed induction of anti-E7 IFNγ-producing cells in the cervix lymphocytes obtained from these patients. E6/E7 siRNA therapy requires a delivery system for its systemic intravenous administration. We here demonstrated that intravenous injection of HPV16 or 18 E6/E7 siRNA polymeric micelles suppressed excellently an increase in size of subcutaneous tumor formed by SiHa or HeLa cell, respectively. Our drug-delivery technology using polymeric micelles enabled the successful systemic administration of siRNA to exhibit anti-tumor effect.
Assuntos
Alphapapillomavirus , Sistemas de Liberação de Medicamentos , Descoberta de Drogas , Terapia de Alvo Molecular , Proteínas Oncogênicas Virais , Proteínas E7 de Papillomavirus , RNA Interferente Pequeno/administração & dosagem , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/virologia , Alphapapillomavirus/imunologia , Colo do Útero , Descoberta de Drogas/tendências , Feminino , Células HeLa , Humanos , Imunoterapia , Cadeias beta de Integrinas , Interferon gama , Lacticaseibacillus casei , Masculino , Micelas , Mucosa/citologia , Vacinas contra Papillomavirus/uso terapêutico , Polímeros , RNA Interferente Pequeno/uso terapêuticoRESUMO
Human papillomavirus (HPV) E6 and E7 oncogenes are essential for the immortalization and maintenance of HPV-associated cancer and are ubiquitously expressed in cervical cancer lesions. Small interfering RNA (siRNA) coding for E6 and E7 oncogenes is a promising approach for precise treatment of cervical cancer, yet a delivery system is required for systemic delivery to solid tumors. Here, an actively targeted polyion complex (PIC) micelle was applied to deliver siRNAs coding for HPV E6/E7 to HPV cervical cancer cell tumors in immune-incompetent tumor-bearing mice. A cell viability assay revealed that both HPV type 16 and 18 E6/E7 siRNAs (si16E6/E7 and si18E6/E7, respectively) interfered with proliferation of cervical cancer cell lines in an HPV type-specific manner. A fluorescence imaging biodistribution analysis further revealed that fluorescence dye-labeled siRNA-loaded PIC micelles efficiently accumulated within the tumor mass after systemic administration. Ultimately, intravenous injection of si16E6/E7 and si18E6/E7-loaded PIC micelles was found to significantly suppress the growth of subcutaneous SiHa and HeLa tumors, respectively. The specific activity of siRNA treatment was confirmed by the observation that p53 protein expression was restored in the tumors excised from the mice treated with si16E6/E7- and si18E6/E7-loaded PIC micelles for SiHa and HeLa tumors, respectively. Therefore, the actively targeted PIC micelle incorporating HPV E6/E7-coding siRNAs demonstrated its therapeutic potential against HPV-associated cancer.
Assuntos
Proteínas de Ligação a DNA/genética , Proteínas Oncogênicas Virais/genética , Proteínas E7 de Papillomavirus/genética , RNA Interferente Pequeno/administração & dosagem , Proteínas Repressoras/genética , Animais , Linhagem Celular Tumoral , Proliferação de Células , Portadores de Fármacos , Feminino , Expressão Gênica , Inativação Gênica , Xenoenxertos , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Camundongos SCID , Micelas , Papillomaviridae , Polietilenoglicóis/química , Polilisina/química , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/terapia , Neoplasias do Colo do Útero/virologiaRESUMO
Interstitial pneumonitis after treatment with pegylated liposomal doxorubicin (PLD) has been rarely reported. We describe herein a case of interstitial pneumonitis in a 49-year-old woman with relapsed ovarian carcinoma treated with PLD. Twenty-five days after the second administration of PLD, she presented with fever and dry cough, and chest CT scans revealed bilateral interstitial infiltrates and ground-glass opacities. She was diagnosed to have interstitial pneumonitis induced by PLD. Steroid therapy improved her symptoms.