RESUMEN
Photodynamic therapy (PDT) using the photosensitizer talaporfin sodium (talaporfin) is a new mode of treatment for cancer. However, the metabolic mechanism of talaporfin has not been clarified. Thus, we investigated the uptake, transportation, and elimination mechanisms of talaporfin in carcinoma and sarcoma. The results showed that talaporfin co-localized in early endosomes and lysosomes. Talaporfin uptake was via clathrin- and caveolae-dependent endocytosis and a high amount of intracellular ATP was essential. Inhibition of lysosomal enzymes maintained intracellular talaporfin levels. Inhibition of K-Ras signaling reduced talaporfin uptake in carcinoma and sarcoma cell lines. Talaporfin was taken up by clathrin- and caveolae-dependent endocytosis, translocated from early endosomes to lysosomes, and finally degraded by lysosomes. We also demonstrated that ATP is essential for the uptake of talaporfin and that activation of K-Ras is involved as a regulatory mechanism. These results provide new insights into the metabolism of talaporfin in cancer cells for the enhancement of PDT for carcinoma and sarcoma.
Asunto(s)
Carcinoma , Fármacos Fotosensibilizantes/metabolismo , Porfirinas/metabolismo , Sarcoma , Línea Celular Tumoral , HumanosRESUMEN
BACKGROUND: Peptide-vaccination therapy targeting tumour-associated antigens can elicit immune responses, but cannot be used to eliminate large tumour burden. In this study, we developed a therapeutic single-chain variable-fragment (scFv) antibody that recognises the cancer stem-like cell/cancer-initiating cell (CSC/CIC) antigen, DNAJB8. METHODS: We screened scFv clones reacting with HLA-A24:20/DNAJB8-derived peptide (DNAJB8_143) complex using naive scFv phage-display libraries. Reactivity and affinity of scFv clones against the cognate antigen were quantified using FACS and surface plasmon resonance. Candidate scFv clones were engineered to human IgG1 (hIgG1) and T-cell-engaging bispecific antibody (CD3xJB8). Complement-dependent cytotoxicity (CDC) and bispecific antibody-dependent cellular cytotoxicity (BADCC) were assessed. RESULTS: scFv clones A10 and B10 were isolated after bio-panning. Both A10-hIgG1 and B10-hIgG1 reacted with DNAJB8-143 peptide-pulsed antigen-presenting cells and HLA-A24(+)/DNAJB8(+) renal cell carcinoma and osteosarcoma cell lines. A10-hIgG1 and B10-hIgG1 showed strong affinity with the cognate HLA/peptide complex (KD = 2.96 × 10-9 M and 5.04 × 10-9 M, respectively). A10-hIgG1 and B10-hIgG1 showed CDC against HLA-A24(+)/DNAJB8(+) cell lines. B10-(CD3xJB8) showed superior BADCC to A10-(CD3xJB8). CONCLUSION: We isolated artificial scFv antibodies reactive to CSC/CIC antigen DNAJB8-derived peptide naturally present on renal cell carcinoma and sarcoma. Immunotherapy using these engineered antibodies could be promising.
Asunto(s)
Antígeno HLA-A24/inmunología , Proteínas del Choque Térmico HSP40/inmunología , Inmunoterapia/métodos , Chaperonas Moleculares/inmunología , Células Madre Neoplásicas/inmunología , Proteínas del Tejido Nervioso/inmunología , Ingeniería de Proteínas/métodos , Anticuerpos de Cadena Única/biosíntesis , Especificidad de Anticuerpos , Citotoxicidad Celular Dependiente de Anticuerpos , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Neoplasias Óseas/inmunología , Neoplasias Óseas/patología , Neoplasias Óseas/terapia , Vacunas contra el Cáncer/biosíntesis , Vacunas contra el Cáncer/uso terapéutico , Carcinoma de Células Renales/inmunología , Carcinoma de Células Renales/patología , Carcinoma de Células Renales/terapia , Línea Celular Tumoral , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/metabolismo , Células HEK293 , Antígeno HLA-A24/genética , Antígeno HLA-A24/metabolismo , Proteínas del Choque Térmico HSP40/genética , Proteínas del Choque Térmico HSP40/metabolismo , Células HT29 , Humanos , Neoplasias Renales/inmunología , Neoplasias Renales/patología , Neoplasias Renales/terapia , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Osteosarcoma/inmunología , Osteosarcoma/patología , Osteosarcoma/terapia , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/metabolismo , Biblioteca de Péptidos , Anticuerpos de Cadena Única/uso terapéuticoAsunto(s)
Hipertensión/complicaciones , Enfermedades Renales/etiología , Enfermedades Renales/terapia , Albuminuria/tratamiento farmacológico , Albuminuria/etiología , Amidas/uso terapéutico , Bloqueadores del Receptor Tipo 1 de Angiotensina II/uso terapéutico , Antihipertensivos/uso terapéutico , Agonistas de los Canales de Calcio/uso terapéutico , Ablación por Catéter , Circulación Cerebrovascular/fisiología , Enfermedad Crónica , Nefropatías Diabéticas/terapia , Quimioterapia Combinada , Medicina Basada en la Evidencia , Fumaratos/uso terapéutico , Humanos , Imidazoles/uso terapéutico , Riñón/inervación , Ensayos Clínicos Controlados Aleatorios como Asunto , Tetrazoles/uso terapéutico , Valina/análogos & derivados , Valina/uso terapéutico , ValsartánRESUMEN
Development and application of anti-CTLA-4 antibody and anti-PD-1 antibody to cancer immunotherapy brought great survival benefits to advanced cancer patients. They have been applied to various cancers such as melanoma, non-small cell lung cancer, renal cell cancer, Hodgkin's disease, and head and neck cancers, and there is no doubt that immunotherapy is becoming a standard therapy as well as surgery, chemotherapy, and radiotherapy. On the other hand, immune-related adverse events (irAEs) have been increasingly reported. Nevertheless, mechanisms of the immune-mediated toxicities are still unclear. There has been a growing interest in the elucidation of the mechanisms. This review describes the general characteristics of irAEs induced by immune checkpoint inhibitors, especially 1. Heterogeneity, 2. Multiplicity, 3. Durability, and 4. Correlativity.