RESUMO
OBJECTIVE: Although many reports describe the short-term hearing outcomes of surgically managed labyrinthine fistulae, the long-term results remain unknown. We reviewed the long-term postoperative hearing outcomes of 14 ears of patients with cholesteatoma and labyrinthine fistulae. METHODS: Between 1996 and 2010, 84 patients with cholesteatoma and labyrinthine fistula underwent tympanoplasty at Hyogo College of Medicine Hospital. Fistulae were located in the lateral semicircular canal in all patients and in the superior semicircular canal in one. Fourteen patients were followed up for more than 5 years. RESULTS: The postoperative air-bone gap was ≤10dB in one patient, between 11 and 20dB in seven, between 21 and 30dB in four, and ≥31dB in two. Mean bone-conduction hearing levels on the operated side had deteriorated by 3, -1 and -2dB at 1, 2 and 4kHz, respectively at 1 year postoperatively, and by 8, 6 and 2dB at 1, 2 and 4kHz, at 5 years postoperatively. Bone-conduction hearing levels at 1 and 2kHz were significantly deteriorated at 5 years postoperatively, compared with baseline and 1 year (P<0.05).
Assuntos
Condução Óssea , Colesteatoma da Orelha Média/cirurgia , Fístula/cirurgia , Doenças do Labirinto/cirurgia , Canais Semicirculares/cirurgia , Timpanoplastia/métodos , Idoso , Estudos de Coortes , Feminino , Perda Auditiva Condutiva , Testes Auditivos , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
BACKGROUND: While the benefit of passive immunotherapy is commonly accepted, active immunization may have advantages for the patient's quality of life. We identified a new epitope of Mab CH401 against Her-2/neu extracellular domain (N: 167-175), and evaluated the effect of active immunization of the 20mer peptide containing the epitope (CH401 peptide). MATERIALS AND METHODS: Epitope-mapping was performed using ELISA with Her-2/neu-related multiple antigen peptides (MAP). BALB/c mice were transplanted with Her-2/neu-expressing lymphoma cell line and immunized with the peptides. For monitoring the condition, ELISA and flow cytometry was performed. RESULTS: CH401 peptide induced Her-2/neu-specific IgG antibody. Tumor growth in immunized mice was suppressed and tumor-infiltrating lymphocytes comprised more CD8(+) T-cells, which secreted larger amounts of interleukin-2 after the peptide re-stimulation. CONCLUSION: The new Her-2/neu peptide contained epitopes for CD4(+) and CD8(+) T-cells, which contributes to the suppressive effect on Her-2/neu-expressing tumor cell growth.
Assuntos
Antineoplásicos/farmacologia , Vacinas Anticâncer/imunologia , Epitopos de Linfócito B/imunologia , Peptídeos/imunologia , Peptídeos/farmacologia , Receptor ErbB-2/imunologia , Vacinação , Motivos de Aminoácidos , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Antígenos de Neoplasias/química , Antígenos de Neoplasias/imunologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Mapeamento de Epitopos , Epitopos de Linfócito B/química , Feminino , Humanos , Depleção Linfocítica , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Transplante de Neoplasias , Peptídeos/química , Receptor ErbB-2/químicaRESUMO
Peptides which mimic functional activities of glycosphingolipids were prepared by a technology of phage-displayed peptide library using monoclonal antibodies against glycosphingolipids. These peptides were named glyco-replica peptides. Peptides prepared with anti-GD1alpha antibody by this technology were found to contain WHW as common motif, and they showed suppressive activity not only on adhesion between hepatic sinusoidal endothelial cells and lymphosarcoma RAW117-H10 cells, but also on metastasis of the tumor cell to the liver and lung. The WHW motif seems to be important to mimic the functional activity of the ganglioside GD1alpha. Next, we prepared GD3-replica peptides using a monoclonal antibody against GD3 (4F6). A peptide, GD3-P4 with highest affinity to 4F6 was used to immunize mice to examine if the mice show their immune response to raise antibodies against GD3. We confirmed the immune response and succeeded in the production of a monoclonal antibody (3D2) against GD3. The monoclonal antibody 3D2 showed specific binding to GD3 on a thin-layer chromatography plate and also melanoma tissues. Interestingly, the amino acid sequence of the CDR regions of light and heavy chains showed high similarity with those of the original GD3 monoclonal antibody (4F6) used for the preparation of GD3-replica peptide. The technology of the phage-displayed peptide library was applied to in vivo bio-panning study using an angiogenesis experimental model. The obtained peptides were found to show strong binding property to the neo-vasculature system and to be quite useful to carry an anti-tumor drug to the tumor tissue. Based on these experimental results, we discuss about some applications of this method to drug discovery.
Assuntos
Desenho de Fármacos , Glicômica/métodos , Biblioteca de Peptídeos , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Adesão Celular/efeitos dos fármacos , Gangliosídeo G(M1)/análogos & derivados , Gangliosídeo G(M1)/farmacologia , Gangliosídeos/imunologia , Glicoesfingolipídeos/química , Glicoesfingolipídeos/metabolismo , Humanos , Melanoma/irrigação sanguínea , Melanoma/imunologia , Melanoma/patologia , Camundongos , Dados de Sequência Molecular , Metástase Neoplásica , Neovascularização Patológica , Peptídeos/química , Peptídeos/imunologia , Peptídeos/farmacologiaRESUMO
This study documents a case of an epithelial-myoepithelial carcinoma (EMC) in the left nasal cavity. A 70-year-old woman who presented with recurrent epistaxis of the left nasal nostril of 3 months duration was found to have a polypoid tumor in the left nasal cavity. A computed tomography (CT) scan revealed a tumor to occupy the left inferior and middle nasal cavity which had destroyed the inferior nasal turbinate, and a horizontal scan showed the tumor to occupy the middle and posterior nasal cavity. Since the tumor was connected to the lateral wall of the left nasal cavity with a narrow stalk, the tumor was excised by peeling the mucosa from the wall of the left nasal cavity. Based on the histological and immunohistochemical findings, the tumor was diagnosed to be an EMC. The follow-up at 12 months after the operation showed no evidence of recurrence.
Assuntos
Mioepitelioma/diagnóstico , Cavidade Nasal , Pólipos Nasais/diagnóstico , Neoplasias Nasais/diagnóstico , Idoso , Biomarcadores Tumorais/análise , Diagnóstico Diferencial , Epistaxe/etiologia , Feminino , Humanos , Mioepitelioma/patologia , Mioepitelioma/cirurgia , Mucosa Nasal/patologia , Mucosa Nasal/cirurgia , Pólipos Nasais/patologia , Pólipos Nasais/cirurgia , Neoplasias Nasais/patologia , Neoplasias Nasais/cirurgia , Recidiva , Conchas Nasais/patologiaRESUMO
GD3-replica peptides were obtained from a phage peptide library and an anti-GD3 monoclonal antibody (Mab) (4F6), and anti-GD3 Mabs were generated by immunizing a peptide GD3P4. A Mab, 3D2 was found to recognize GD3 by immunohistochemical approaches. Amino acid analysis of heavy and light chain variable regions of 4F6 and 3D2 showed that the respective chains had the same length, and only a few different amino acid substitutions were found. The present data indicate that the immunogenic GD3P4 is processed in a certain size and exposed on the antigen-presenting cells with a molecular shape quite similar to that of the GD3 epitope in 4F6.
Assuntos
Anticorpos Monoclonais/química , Formação de Anticorpos , Gangliosídeos/imunologia , Peptídeos/imunologia , Aminoácidos/análise , Animais , Anticorpos Monoclonais/biossíntese , Apresentação de Antígeno , Sequência de Bases , Imunização , Cadeias Pesadas de Imunoglobulinas , Cadeias Leves de Imunoglobulina , Região Variável de Imunoglobulina , Camundongos , Mimetismo Molecular , Dados de Sequência Molecular , Biblioteca de PeptídeosRESUMO
Because membrane type-1 matrix metalloproteinase (MT1-MMP) is expressed specifically on the angiogenic endothelium as well as tumor cells, an agent possessing the ability to bind to this molecule might be useful as a tool for active targeting of tumor angiogenic vessels. Based on the sequences of peptide substrates of MT1-MMP, which had been determined by using a phage-displayed peptide library, we examined the binding ability of peptide-modified liposomes for endothelial cells and targeting ability for tumor tissues by positron emission tomography (PET). Liposomes modified with stearoyl-Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR-Lip) showed high binding ability to human umbilical vein endothelial cells and accumulated in the tumor about 4-fold more than did the unmodified liposomes. Because we reported previously that liposomalized 5'-O-dipalmitoylphosphatidyl 2'-C-cyano-2'-deoxy-1-beta-D-arabino-pentofuranosylcytosine (DPP-CNDAC), a hydrophobized derivative of the novel antitumor nucleoside CNDAC, strongly suppressed tumor growth when delivered in liposomes modified with another angiogenic homing peptide, we examined the antitumor activity of DPP-CNDAC entrapped in GPLPLR-Lip. DPP-CNDAC/GPLPLR-Lip showed significant tumor growth suppression compared to DPP-CNDAC/unmodified liposomes. These results suggest that DPP-CNDAC-liposomes modified with MT1-MMP-targeted peptide are useful for cancer anti-neovascular therapy (ANET), namely, tumor growth suppression by damage to angiogenic endothelial cells.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Arabinonucleotídeos/uso terapêutico , Neoplasias do Colo/irrigação sanguínea , Endotélio Vascular/metabolismo , Metaloendopeptidases/antagonistas & inibidores , Neovascularização Patológica/tratamento farmacológico , Animais , Protocolos de Quimioterapia Combinada Antineoplásica , Neoplasias do Colo/enzimologia , Sistemas de Liberação de Medicamentos , Endotélio Vascular/enzimologia , Endotélio Vascular/patologia , Humanos , Lipossomos , Masculino , Metaloproteinase 14 da Matriz , Metaloproteinases da Matriz Associadas à Membrana , Camundongos , Camundongos Endogâmicos BALB C , Neovascularização Patológica/enzimologia , Ácidos Fosfatídicos/química , Ácidos Fosfatídicos/metabolismo , Transporte Proteico , Taxa de Sobrevida , Resultado do Tratamento , Células Tumorais Cultivadas/transplante , Veias Umbilicais/metabolismo , Veias Umbilicais/patologiaAssuntos
Inibidores da Angiogênese/uso terapêutico , Antineoplásicos/uso terapêutico , Arabinonucleotídeos/administração & dosagem , Neovascularização Patológica , Fotoquimioterapia/métodos , Porfirinas/administração & dosagem , Radiossensibilizantes/administração & dosagem , Inibidores da Angiogênese/administração & dosagem , Animais , Antineoplásicos/administração & dosagem , Arabinonucleotídeos/uso terapêutico , Portadores de Fármacos , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Humanos , Lipossomos , Camundongos , Oligopeptídeos , Porfirinas/uso terapêutico , Radiossensibilizantes/uso terapêutico , Sarcoma/tratamento farmacológicoRESUMO
Novel peptides homing to angiogenic vessels were recently isolated from a phage-displayed random pentade-capeptide library, and peptides having WRP sequence showed tumor growth suppression. In this study, we observed that another novel sequence, PVVLFPLH, suppressed tumor growth in vivo. Through the study of tumor growth suppression by the 5-mer peptides derived from this sequence, we determined the epitope sequence to be LFPLH. LFPLH, but not the shuffled peptide FHLLP, suppressed the migration of vascular endothelial growth factor-stimulated human umbilical vein endothelial cells. Interestingly, growth suppression of LFPLH against the cells as well as tumor cells was not observed in vitro. Therefore LFPLH may function to induce tumor dormancy through inhibition of angiogenesis.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Neovascularização Patológica/tratamento farmacológico , Peptídeos/uso terapêutico , Sarcoma Experimental/tratamento farmacológico , Inibidores da Angiogênese/química , Animais , Divisão Celular/efeitos dos fármacos , Endotélio Vascular/citologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/uso terapêutico , Biblioteca de Peptídeos , Peptídeos/química , Sarcoma Experimental/irrigação sanguínea , Veias Umbilicais/citologiaRESUMO
We previously reported that liposomalized 5'-O-dipalmitoylphosphatidyl 2'-C-cyano-2'-deoxy-1-beta-D-arabino-pentofuranosylcytosine (DPP-CNDAC), a hydrophobized derivative of the novel antitumor nucleoside CNDAC, is quite useful for cancer therapy. On the other hand, for anti-neovascular therapy, we recently isolated peptides homing to angiogenic vessels from a phage-displayed random peptide library, and observed that peptide-modified liposomal adriamycin strongly suppressed tumor growth, perhaps through damaging angiogenic endothelial cells. In the present study, we modified DPP-CNDAC-liposomes with one of the angiogenic homing peptides, APRPG, and examined their antitumor activity. Three doses of APRPG-modified DPP-CNDAC-liposomes (15 mg/kg as CNDAC) strongly inhibited tumor growth compared with the same number of doses of unmodified DPP-CNDAC-liposomes. The life span was increased 31.8%, with one completely cured mouse out of the six mice treated. Since the accumulation of liposomes in the tumor tissue was not so much different between APRPG-liposomes and non-modified liposomes, the enhanced therapeutic efficacy may be explained as the alteration of targets, i.e. APRPG-modified DPP-CNDAC-liposomes caused tumor growth suppression through damage of angiogenic endothelial cells. Anti-neovascular therapy promises no drug resistance, and should be effective against essentially any kind of solid tumor; and thus the present results demonstrate another benefit of the therapy, namely, high efficacy of cancer treatment.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Citarabina/análogos & derivados , Citarabina/uso terapêutico , Neoplasias Experimentais/prevenção & controle , Neovascularização Patológica/prevenção & controle , Ácidos Fosfatídicos/uso terapêutico , Sequência de Aminoácidos , Inibidores da Angiogênese/farmacocinética , Animais , Citarabina/química , Citarabina/farmacocinética , Sistemas de Liberação de Medicamentos , Lipossomos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Neoplasias Experimentais/irrigação sanguínea , Neovascularização Patológica/patologia , Oligopeptídeos/química , Oligopeptídeos/farmacocinética , Oligopeptídeos/uso terapêutico , Ácidos Fosfatídicos/química , Ácidos Fosfatídicos/farmacocinética , Análise de Sobrevida , Fatores de Tempo , Distribuição Tecidual , Resultado do Tratamento , Células Tumorais CultivadasRESUMO
Cancer chemotherapy targeted to angiogenic vessels is expected to cause indirect tumor regression through the damage of the neovasculature without the induction of drug resistance. To develop a tool for neovasculature-specific drug delivery, we isolated novel peptides homing to angiogenic vessels formed by a dorsal air sac method from a phage-displayed peptide library. Three distinct phage clones that markedly accumulated in murine tumor xenografts presented PRPGAPLAGSWPGTS-, DRWRPALPVVLFPLH- or ASSSYPLIHWRPWAR-peptide respectively. After the determination of the epitope sequences of these peptides, we modified liposomes with epitope penta-peptides. Liposome modified with APRPG-peptide showed high accumulation in murine tumor xenografts, and APRPG-modified liposome encapsulating adriamycin effectively suppressed experimental tumor growth. Finally, specific binding of APRPG-modified liposome to human umbilical endothelial cells, and that of PRP-containing peptide to angiogenic vessels in human tumors, i.e., islet cell tumor and glioblastoma, were demonstrated. The present study indicates the usefulness of APRPG-peptide as a tool for anti-neovascular therapy, a novel modality of cancer treatment.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Melanoma Experimental/irrigação sanguínea , Neovascularização Patológica/tratamento farmacológico , Peptídeos/uso terapêutico , Sarcoma Experimental/irrigação sanguínea , Sequência de Aminoácidos , Animais , Antibióticos Antineoplásicos/farmacologia , Divisão Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Doxorrubicina/farmacologia , Endotélio Vascular/efeitos dos fármacos , Humanos , Injeções Subcutâneas , Lipossomos/metabolismo , Linfocinas/farmacologia , Masculino , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Confocal , Dados de Sequência Molecular , Neovascularização Patológica/patologia , Biblioteca de Peptídeos , Sarcoma Experimental/patologia , Tomografia Computadorizada de EmissãoRESUMO
Novel peptides homing to angiogenic vessels were recently isolated from a phage-displayed random pentadecapeptide library. One of the isolated peptides, ASSSYPLIHWRPWAR, significantly suppressed the migration of VEGF-stimulated human umbilical vein endothelial cells. Dendoric ASSSYPLIHWRPWAR-peptide suppressed the formation of new blood vessels in dorsal air sac model mice. Furthermore, ASSSYPLIHWRPWAR-peptide and the fragment peptides containing WRP, which is revealed to be an epitope sequence, significantly suppressed the tumor growth, although 15-mer shuffled peptide derived from ASSSYPLIHWRPWAR and pentapeptides with alanine substitution of each residue of WRP did not. Taken together, ASSSYPLIHWRPWAR-peptide may cause tumor dormancy through inhibition of angiogenesis, and the WRP sequence may be the minimal and essential sequence for this activity.