Asunto(s)
Micosis Fungoide , Parapsoriasis , Receptores de Antígenos de Linfocitos T , Neoplasias Cutáneas , Humanos , Micosis Fungoide/genética , Micosis Fungoide/patología , Micosis Fungoide/inmunología , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/inmunología , Parapsoriasis/genética , Parapsoriasis/patología , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/inmunología , Masculino , Femenino , Persona de Mediana Edad , Factores de TiempoRESUMEN
Melanoma is one of the most severe skin cancers, derived from melanocytes. Among various therapies for melanoma, adoptive immunotherapy using tumor-infiltrating lymphocytes/chimeric antigen receptor-T cells (TCs) is advanced in recent years; however, the efficacy is still limited, and major challenges remain in terms of safety and cell supply. To solve the issues of adoptive immunotherapy, we utilized induced pluripotent stem cells (iPSCs), which have an unlimited proliferative ability and various differentiation capability. First, we monoclonally isolated CD8+ TCs specifically reactive with NY-ESO-1, one of tumor antigens, from the melanoma patient's monocytes after stimulated with NY-ESO-1 peptide by manual procedure, and cultured NY-ESO-1-specific TCs until proliferated and formed colonies. iPSCs were consequently generated from colony-forming TCs by exogenous expression of reprogramming factors using Sendai virus vector. After the RAG2 gene in TC-derived iPSCs (T-iPSCs) was knocked out for preventing T-cell receptor (TCR) rearrangement, T-iPSCs were re-differentiated into rejuvenated cytotoxic TCs. We confirmed that TCR of T-iPSC-derived TC was maintained as the same of original TCs. In conclusion, T-iPSCs have a potential to be an unlimited cell source for providing cytotoxic TCs. Our study could be a "touchstone" to develop iPSC-based adoptive immunotherapy for the treatment of melanoma for the future clinical use.
Asunto(s)
Células Madre Pluripotentes Inducidas , Melanoma , Humanos , Linfocitos T Citotóxicos/metabolismo , Inmunoterapia Adoptiva , Proyectos Piloto , Células Madre Pluripotentes Inducidas/metabolismo , Melanoma/patología , Linfocitos T CD8-positivos/metabolismo , Receptores de Antígenos de Linfocitos T/metabolismo , Antígenos de Neoplasias , InmunoterapiaRESUMEN
Malignant melanoma (MM) is one of the most aggressive, recalcitrant, and recurrence-prone skin neoplasms. Its feature is likely to be associated with phenotypic conversion due to tumor heterogeneity. The multidisciplinary assessment, including surgery, drug therapy using anticancer agents and immune checkpoint inhibitors, and radiotherapy, is needed for the treatment of advanced MM. Herein, we report a long-term follow-up MM, in which multiple phenotypic conversion occurred during several treatments. In particular, our case obtained granulocyte colony-stimulating factor-producing ability during the intermission of nivolumab therapy and it was successfully controlled by re-administration of nivolumab. Sharing the case having a varied clinical course is meaningful to increase the knowledge and decision branches for the treatment of melanoma.
RESUMEN
CD30-positive large cell transformation that occurs in early mycosis fungoides potentially possesses characteristics of spontaneous regression as with CD30-positive lymphoproliferative disorders. Such transformation may not relate to poor prognosis.
RESUMEN
BACKGROUND: Recessive dystrophic epidermolysis bullosa (RDEB) is a monogenic skin blistering disorder caused by mutations in the type VII collagen gene. A combination of biological technologies, including induced pluripotent stem cells (iPSCs) and several gene-editing tools, allows us to develop gene and cell therapies for such inherited diseases. However, the methodologies for gene and cell therapies must be continuously innovated for safe clinical use. OBJECTIVE: In this study, we used the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology to correct the pathogenic mutation in RDEB-specific iPSCs, and the piggyBac transposon system so that no residual gene fragments remained in the genome of iPSCs after correcting the mutation. METHODS: For homologous recombination (HR)-based gene editing using CRISPR/Cas9, we designed guide RNA and template DNA including homologous sequences with drug-mediated selection cassette flanked by inverted repeat sequences of the transposon. HR reaction using CRISPR/Cas9 was induced in RDEB-specific iPSCs, and mutation-corrected iPSCs (MC-iPSCs) was obtained. Consequently, the selection cassette in the genome of MC-iPSCs was removed by transposase expression. RESULTS: After CRISPR/Cas9-induced gene editing, we confirmed that the pathogenic mutation in RDEB-specific iPSCs was properly corrected. In addition, MC-iPSCs had no genetic footprint after removing the selection cassette by transposon system, and maintained their "stemness". When differentiating MC-iPSCs into keratinocytes, the expression of type VII collagen was restored. CONCLUSIONS: Our study demonstrated one of the safer approaches to establish gene and cell therapies for skin hereditary disorders for future clinical use.
Asunto(s)
Colágeno Tipo VII/genética , Epidermólisis Ampollosa Distrófica/terapia , Edición Génica/métodos , Células Madre Pluripotentes Inducidas/metabolismo , Queratinocitos/trasplante , Sistemas CRISPR-Cas/genética , Diferenciación Celular , Línea Celular , Colágeno Tipo VII/metabolismo , Elementos Transponibles de ADN/genética , Epidermólisis Ampollosa Distrófica/genética , Terapia Genética/métodos , Recombinación Homóloga , Humanos , Queratinocitos/metabolismo , MutaciónRESUMEN
BACKGROUND: Sebaceous carcinoma is a rare but progressive malignant skin cancer, and the incidence is approximately five times higher in post-transplant patients than in people who have not received kidney transplants. Sebaceous carcinoma is sometimes found concurrently with visceral cancers and a genetic abnormality, Muir-Torre syndrome. We report the case of a female kidney transplant recipient with sebaceous carcinoma concurrent with colon cancer 10 years after transplantation. CASE PRESENTATION: A 43-year-old woman was admitted due to a rapidly progressive tumor on her head. Histologically, the tumor was diagnosed as sebaceous carcinoma. We diagnosed her with Muir-Torre syndrome based on the following evidence: 1) high prevalence of microsatellite instability in gene locus assay, 2) absence of mismatch repair proteins in the sebaceous carcinoma on immunohistochemical analysis, and 3) a genetic mutation of 1226_1227delAG in the MSH2 exon 7 in the lesion detected by DNA sequencing analysis. Several reports have shown an association between immunosuppressive agents and latent Muir-Torre syndrome progression. Therefore, the progression of colon cancer in this case originated from her genetic mutation for Muir-Torre syndrome and long-term use of immunosuppressive agents. CONCLUSION: This case report not only highlights the importance of adequate diagnosis and therapy for Muir-Torre syndrome, but also suggests the further prevention of the development of malignant tumors in kidney transplant recipients. Physicians should be mindful that sebaceous carcinoma in kidney transplant recipients is highly concurrent with Muir-Torre syndrome.
Asunto(s)
Adenocarcinoma/genética , Neoplasias del Colon/genética , Neoplasias de Cabeza y Cuello/genética , Trasplante de Riñón/efectos adversos , Síndrome de Muir-Torre/genética , Neoplasias Cutáneas/genética , Adenocarcinoma/patología , Adulto , Neoplasias del Colon/patología , Proteínas de Unión al ADN/análisis , Femenino , Neoplasias de Cabeza y Cuello/patología , Humanos , Inmunosupresores/efectos adversos , Inestabilidad de Microsatélites , Síndrome de Muir-Torre/patología , Proteína 2 Homóloga a MutS/genética , Mutación , Cuero Cabelludo , Neoplasias Cutáneas/patología , Factores de Tiempo , Receptores de TrasplantesRESUMEN
BACKGROUND: Type 1 interferons (IFNs), including IFN-ß, are widely used in adjuvant therapy for patients who undergo surgery for malignant melanoma to inhibit recurrence and in-transit metastasis. The precise mechanisms underlying the tumor-suppressive effects of IFN-ß on melanoma are not yet completely understood. OBJECTIVE: The purpose was to reveal the mechanisms underlying the tumor-suppressive effects of IFN-ß via interleukin (IL)-24. METHODS: Genome-wide oligonucleotide microarray, quantitative real-time reverse transcription-polymerase chain reaction (PCR), enzyme-linked immunosorbent assay and western blotting assay were performed using four melanoma cell lines (A375, RPMI-7951, SK-MEL-5 and SK-MEL-1) treated with natural-type IFN-ß to assess the expression of IL-24. Proliferation assay was performed using these melanoma cells and IL-24 knock-down melanoma cells. RESULTS: Genome-wide microarray analysis detected candidate genes upregulated in IFN-ß-sensitive cells after treatment with IFN-ß. We focused on IL-24 among the candidate genes encoding secretory proteins. Peak IL-24 mRNA expression completely correlated with the order of sensitivity of melanoma cells to IFN-ß. IFN-ß treatment induced extracellular IL-24 protein in IFN-ß-sensitive cells, but did not induce intracellular IL-24 protein. Knock-down of IL-24 changed melanoma cells into IFN-ß-resistant cells. The expression ratio of IL-22R1, one of the IL-24 receptors, correlated with the order of sensitivity of melanoma cells to IFN-ß. Treatment with recombinant human IL-24 did not have any effects on all the melanoma cell lines. CONCLUSION: Our data suggest that IFN-ß suppresses the proliferation of melanoma cells through extracellular IL-24 protein derived from melanoma cells.
Asunto(s)
Interferón beta/administración & dosificación , Interleucinas/administración & dosificación , Melanoma/tratamiento farmacológico , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Quimioterapia Adyuvante , Estudio de Asociación del Genoma Completo , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Receptores de Interleucina/metabolismo , Proteínas Recombinantes/administración & dosificaciónAsunto(s)
Antineoplásicos Inmunológicos/efectos adversos , Linfoma Folicular/tratamiento farmacológico , Rituximab/efectos adversos , Vasculitis Leucocitoclástica Cutánea/inducido químicamente , Complejo Antígeno-Anticuerpo/inmunología , Antígenos CD20/inmunología , Antígenos CD20/metabolismo , Antineoplásicos Inmunológicos/farmacología , Biopsia , Dermis/patología , Humanos , Linfoma Folicular/inmunología , Masculino , Persona de Mediana Edad , Rituximab/farmacología , Vasculitis Leucocitoclástica Cutánea/inmunología , Vasculitis Leucocitoclástica Cutánea/patologíaAsunto(s)
Antifúngicos/efectos adversos , Queratosis Actínica/diagnóstico , Trastornos por Fotosensibilidad/diagnóstico , Rayos Ultravioleta/efectos adversos , Voriconazol/efectos adversos , Anciano , Aspergilosis/tratamiento farmacológico , Aspergilosis/microbiología , Humanos , Queratosis Actínica/etiología , Queratosis Actínica/patología , Masculino , Enfermedades de los Senos Paranasales/tratamiento farmacológico , Enfermedades de los Senos Paranasales/microbiología , Trastornos por Fotosensibilidad/etiología , Trastornos por Fotosensibilidad/patología , Piel/efectos de los fármacos , Piel/patología , Piel/efectos de la radiación , Pruebas CutáneasAsunto(s)
Conducto Auditivo Externo/patología , Pérdida Auditiva Conductiva/tratamiento farmacológico , Hidroxicloroquina/uso terapéutico , Lupus Eritematoso Discoide/complicaciones , Piel/patología , Administración Oral , Anciano , Audiometría , Biopsia , Femenino , Pérdida Auditiva Conductiva/diagnóstico , Pérdida Auditiva Conductiva/etiología , Humanos , Lupus Eritematoso Discoide/tratamiento farmacológico , Lupus Eritematoso Discoide/patología , Resultado del TratamientoAsunto(s)
Receptores de Activinas Tipo II/genética , Telangiectasia Hemorrágica Hereditaria/genética , Análisis Mutacional de ADN , Endoglina/genética , Femenino , Humanos , Imagen por Resonancia Magnética , Microvasos/patología , Persona de Mediana Edad , Mutación , Linaje , Piel/irrigación sanguínea , Piel/patología , Telangiectasia Hemorrágica Hereditaria/diagnóstico por imagen , Telangiectasia Hemorrágica Hereditaria/patologíaAsunto(s)
Folículo Piloso/metabolismo , Complejo Represivo Polycomb 1/genética , Telomerasa/metabolismo , Vibrisas/metabolismo , Animales , Células Cultivadas , Folículo Piloso/citología , Folículo Piloso/crecimiento & desarrollo , Humanos , Ratones , Fenotipo , Complejo Represivo Polycomb 1/metabolismo , Transducción de Señal , Telomerasa/genética , Factores de Tiempo , Transfección , Regulación hacia Arriba , Vibrisas/citologíaRESUMEN
Induced pluripotent stem (iPS) cells have the ability to differentiate into multiple cell types in the body and have an unlimited growth potential. However, iPS cell-derived melanocytes produced by existing protocols have significant limitations in developing novel strategies for regenerative medicine and cell therapies of pigmentation disorders in humans because they involve culture in media containing fetal bovine serum and nonphysiological agents. In this study, we established an in vitro approach to generate iPS cell-derived human melanocytes that have higher proliferation rates and increased melanin production compared with melanocytes prepared by previously reported approaches. Importantly, our iPS cell-derived human melanocytes are prepared in fetal bovine serum-free culture conditions that do not contain any nonphysiological agents. We designed two original methods, transferring black colonies by pipette and recovering black cell pellets from centrifuged medium, and numerous human iPS cell-derived melanocytes proliferated in gelatinous dishes coated with Matrigel after 12 days. We also succeeded in inducing melanin pigmentation in the nude mouse skin in vivo using those human iPS cell-derived melanocytes. We propose that this method using iPS cells established from T cells in the blood of normal human volunteers could be applied clinically to develop regenerative medicine and cell therapies for various forms of human pigmentation disorders.