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Introduction - Hypopharyngeal squamous cell carcinoma has poor prognosis with only 25% chances of 5 years of relative survival in such patients in spite of conventional treatment including radical surgery, radiotherapy and concurrent chemotherapy. Case Presentation - A chronic tobacco - betel nut chewer 62-years-old male patient had dysphagia with hoarseness of voice diagnosed with stage III, grade II malignant pyriform fossa. The patient underwent 9 cycles of neo adjuvant chemotherapy with Inj Paclitaxel 100 mg and Inj Cisplatin 40 mg. He was then referred to our institute for Radical Radiotherapy with Concurrent Chemotherapy with adjunct Ayurvedic treatment. A total dose of 70 Gy of radiation with cobalt 60 source was administered to the bilateral face and neck, in 35 fractions. Patient also received 6 cycles of concurrent weekly chemotherapy with Inj Cisplatin 40 mg. He received well planned adjunct Ayurvedic treatment in the form of Oral Ayurvedic Medicines (OAM) and Detoxifying treatment, Panchakarma. All the measured adverse effects of radiotherapy such as Stomatitis, Xerostomia, Taste Alteration, Dysphagia, Nausea etc. were observed to be remarkably low during and post radiotherapy in this patient. Karnofsky and QoL scores revealed patient's wellbeing throughout the treatment course. After 5 years PET CT scan revealed no FDG avid loco regional recurrence or distant organ involvement implying Disease Free Survival (DFS). Various Chemokines, cytokines and oxidative stress were assessed during the course of treatment to observe tumour microenvironment. Conclusion - Present case of HNC, Stage III and Grade II belonged to high-grade, high-risk hypopharyngeal cancer with poor prognosis. The patient opted for Ayurvedic treatment besides radiotherapy which continued thereafter for 5 years. We therefore emphasize that in this case, minimum side effects of radiotherapy, immunomodulation and reduction in inflammation and oxidative stress along with good quality of life can be attributed to OAM and repeated detoxifying Panchakarma treatment supported with healthy diet and good lifestyle. The highlight of the study is the marked effect on the patient's immune response and reduction in oxidative stress leading to 5 years and beyond of DFS.
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Recent global health concern motivated the exploration of natural medicinal plant resources as an alternative target for treating COVID-19 infection and associated inflammation. In the current study, a phytochemical, 6-shogaol [1-(4-hydroxy-3-methoxyphenyl)dec-4-en-3-one; 6-SHO] was investigated as a potential anti-inflammatory and anti-COVID-19 agent. In virus release assay, 6-SHO efficiently (94.5%) inhibited SARS-CoV2 replication. When tested in the inflammasome activation model, 6-SHO displayed mechanistic action by regulating the expression of the inflammasome pathway molecules. In comparison to the existing drugs, remdesivir and hydroxy-chloroquine, 6-SHO was not only found to be as effective as the standard anti-viral drugs but also much superior and safe in terms of predicted physicochemical properties and clinical toxicity. Comparative molecular dynamics simulation demonstrated a stable interaction of 6-SHO with NLRP3 (the key inflammasome regulator) in the explicit water environment. Overall, this study provides important cues for further development of 6-SHO as potential anti-inflammatory and anti-viral therapeutic agents.
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A library of new phenstatin based indole linked chalcone compounds (9a-z and 9aa-ad) were designed and synthesized. Of these, compound 9a with 1-methyl, 2- and 3-methoxy substituents in the aromatic ring was efficacious against the human oral cancer cell line SCC-29B, spheroids, and in a mouse xenograft model of oral cancer AW13516. Compound 9a exhibited anti-cancer activity through disrupting cellular integrity and affecting glucose metabolism-which is a hallmark of cancer. The cellular architecture was affected by inhibition of tubulin polymerization as observed by an immunofluorescence assay on 9a-treated SCC-29B cells. An in vitro tubulin polymerization kinetics assay provided evidence of direct interaction of 9a with tubulin. This physical interaction between tubulin and compound 9a was further confirmed by Surface Plasmon Resonance (SPR) analysis. Molecular docking experiments and validations revealed that compound 9a interacts and binds at the colchicine binding site of tubulin and at active sites of key enzymes in the glucose metabolism pathway. Based on in silico modeling, biophysical interactions, and pre-clinical observations, 9a consisting of phenstatin based indole-chalcone scaffolds, can be considered as an attractive tubulin polymerization inhibitor candidate for developing anti-cancer therapeutics.
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Antineoplásicos/síntesis química , Benzofenonas/química , Chalcona/síntesis química , Indoles/química , Neoplasias de la Boca/tratamiento farmacológico , Moduladores de Tubulina/síntesis química , Animales , Antineoplásicos/farmacología , Dominio Catalítico , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Chalcona/farmacología , Colchicina/química , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Masculino , Ratones , Simulación del Acoplamiento Molecular , Estructura Molecular , Neoplasias de la Boca/diagnóstico por imagen , Neoplasias Experimentales , Tomografía de Emisión de Positrones , Unión Proteica , Tubulina (Proteína)/metabolismo , Moduladores de Tubulina/farmacologíaRESUMEN
Oral tumor microenvironment is characterized by chronic inflammation signified with infiltrating leukocytes and soluble mediators which cause immune suppression. However, how immunosuppressive cells like myeloid-derived suppressor cells (MDSCs) maintain the immunosuppressive tumor microenvironment and influence T cell function in oral squamous cell carcinoma (OSCC) patients remains poorly understood. In the present study, we found that percentages of MDSCs were higher in oral cancer patients compared to healthy individuals and correlated with cancer stage. Monocytic MDSCs (M-MDSCs) were prevalent in the periphery, while granulocytic/polymorphonuclear subset dominated the tumor compartment. M-MDSCs suppressed the lymphocyte proliferation and decreased the CD3-ζ (zeta) chain expression and interferon gamma production. The percentage of M-MDSCs in peripheral blood correlated inversely with CD3-ζ chain expression in T cells of these patients. Interleukin 6 (IL-6)-induced phosphorylated STAT3-regulated programmed cell death ligand 1, CCAAT/enhancer-binding proteins alpha and beta and Interleukin 10 expression in MDSCs. MDSCs inhibited TGF-ß-driven generation of induced regulatory T cells in vitro. M-MDSCs secreted interleukins IL-6, IL-1ß, IL-23 and PGE2 and facilitated T-helper 17 (Th17) cell differentiation which utilizes nitric oxide synthase and cyclooxygenase 2 enzyme activity. Interestingly, OSCC patients showed increased levels of Th17 cells in peripheral blood and tumor tissue. Thus, increased frequency of MDSCs, Th17 cells and decreased expression of CD3-ζ chain portray T cell tolerance and chronic inflammatory state facilitating tumor growth.
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Carcinoma de Células Escamosas/genética , Neoplasias de la Boca/genética , Células Supresoras de Origen Mieloide/inmunología , Células Th17/inmunología , Diferenciación Celular , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Decreased expression of CD3-ζ chain, an adaptor protein associated with T-cell signalling, is well documented in patients with oral cancer, but the mechanistic justifications are fragmentary. Previous studies in patients with oral cancer have shown that decreased expression of CD3-ζ chain was associated with decreased responsiveness of T cells. Tumours are known to induce localized as well as systemic immune suppression. This study provides evidence that oral tumour-derived factors promote immune suppression by down-regulating CD3-ζ chain expression. 2'5'-Oligoadenylate synthetase 2 (OAS2) was identified by the proteomic approach and our results established a causative link between CD3-ζ chain down-regulation and OAS2 stimulation. The surrogate situation was established by over-expressing OAS2 in a HEK293 cell line and cell-free supernatant was collected. These supernatants when incubated with T cells resulted in down-regulation of CD3-ζ chain, which shows that the secreted OAS2 is capable of regulating CD3-ζ chain expression. Incubation of T cells with cell-free supernatants of oral tumours or recombinant human OAS2 (rh-OAS2) induced caspase-3 activation, which resulted in CD3-ζ chain down-regulation. Caspase-3 inhibition/down-regulation using pharmacological inhibitor or small interfering RNA restored down-regulated CD3-ζ chain expression in T cells induced by cell-free tumour supernatant or rh-OAS2. Collectively these results show that OAS2 leads to impairment in CD3-ζ chain expression, so offering an explanation that might be applicable to the CD3-ζ chain deficiency observed in cancer and diverse disease conditions.
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2',5'-Oligoadenilato Sintetasa/metabolismo , Complejo CD3/metabolismo , Caspasa 3/metabolismo , Linfocitos Infiltrantes de Tumor/enzimología , Neoplasias de la Boca/enzimología , Linfocitos T/enzimología , 2',5'-Oligoadenilato Sintetasa/genética , Complejo CD3/inmunología , Estudios de Casos y Controles , Caspasa 3/genética , Línea Celular Tumoral , Regulación hacia Abajo , Activación Enzimática , Regulación Neoplásica de la Expresión Génica , Células HEK293 , Humanos , Linfocitos Infiltrantes de Tumor/inmunología , Neoplasias de la Boca/genética , Neoplasias de la Boca/inmunología , Neoplasias de la Boca/patología , Comunicación Paracrina , Proteómica/métodos , Interferencia de ARN , Proteínas Recombinantes/metabolismo , Transducción de Señal , Linfocitos T/inmunología , Factores de Tiempo , Transfección , Células Tumorales CultivadasRESUMEN
Immune dysfunction is the hallmark of patients with oral cancer. Down-regulation of T cell receptor (TCR) zeta chain expression was observed in T cells from patients with oral squamous cell carcinoma. In peripheral blood, the decrease in TCR zeta chain showed an inverse correlation with the tumor stage as demonstrated by western blotting, confocal microscopy and flow cytometry. The mechanism of TCR zeta chain degradation in the peripheral blood involves ubiquitination and subsequent targeting of TCR zeta for degradation in the lysosome. Decreased expression of PKC theta and the subsequent decrease of TCR zeta chain transcription factor Elf-1 and its binding to DNA may contribute to the decreased/or absent TCR zeta chain transcripts in the tumor infiltrating lymphocytes. Oral cancer patients exhibiting TCR zeta chain defect also showed impaired lymphocyte proliferation, cytokine profile and intracellular calcium release upon stimulation with anti CD3 mAb. Our data shows that posttranslational degradation is primarily responsible for decreased TCR zeta chain expression in the peripheral blood, while a transcriptional defect is observed in the tumor compartment. The down-regulation of TCR zeta chain culminates into impaired lymphocyte responses in these patients.
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Carcinoma de Células Escamosas/inmunología , Neoplasias de la Boca/inmunología , Receptores de Antígenos de Linfocitos T/metabolismo , Western Blotting , Carcinoma de Células Escamosas/sangre , Carcinoma de Células Escamosas/patología , Estudios Transversales , Regulación hacia Abajo , Ensayo de Cambio de Movilidad Electroforética , Ensayo de Inmunoadsorción Enzimática , Efrina-B2/metabolismo , Citometría de Flujo , Regulación Neoplásica de la Expresión Génica , Humanos , Microscopía Confocal , Neoplasias de la Boca/sangre , Neoplasias de la Boca/patología , Estadificación de Neoplasias , Receptores de Antígenos de Linfocitos T/inmunología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Transcripción GenéticaRESUMEN
We have analyzed TCR Vbeta gene usage and clonality of T cells in the peripheral blood of patients with oral cancer and healthy individuals. A large repertoire of clonal TCR Vbeta was observed in the peripheral blood of cancer patients, and this clonal expansion was dominantly represented in the CD8+ T cells. A marked decrease in the lymphocyte proliferative responses to mitogen and anti-CD3 MAb and spontaneous apoptosis was observed in lymphocytes. Appropriate co-stimulation of lymphocytes (anti-TCR Vbeta MAb and anti-CD28 MAb) restored the lymphocyte proliferative responses and CD3-zeta chain expression in these patients.
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Linfocitos T CD8-positivos/inmunología , Activación de Linfocitos , Neoplasias de la Boca/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/sangre , Adulto , Anticuerpos Monoclonales , Apoptosis , Antígenos CD28/sangre , Complejo CD3/sangre , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/patología , Estudios de Casos y Controles , Proliferación Celular , Células Cultivadas , Células Clonales/inmunología , Femenino , Regulación Neoplásica de la Expresión Génica , Genotipo , Humanos , Activación de Linfocitos/efectos de los fármacos , Masculino , Persona de Mediana Edad , Mitógenos/farmacología , Neoplasias de la Boca/genética , Neoplasias de la Boca/patología , Fenotipo , Receptores de Antígenos de Linfocitos T alfa-beta/genéticaRESUMEN
We analyzed the T cell receptor (TCR) gammadelta gene repertoire in peripheral blood and tumor compartment of oral cancer (OC) patients before and after stimulation with heat shock proteins (hsp), which are known ligands for gammadelta T cells. Clonal TCR gamma and delta gene rearrangements in lymphocytes from tumor compartment and peripheral blood were studied using TCR Vgamma and Vdelta gene primers in PCR followed by heteroduplex analysis. Vgamma gene segments derived from VgammaI or VgammaII gene families were most dominantly expressed in peripheral blood lymphocytes (PBL) as compared to tumor infiltrating lymphocytes (TIL) of OC patients. Of the rearranged TCR delta alleles Vdelta1-Jdelta1 and Vdelta2-Jdelta1 gene rearrangements were the most predominant in PBL and TIL of OC patients respectively. Stimulation of gammadelta T cells with hsp 60/70 demonstrated a selective clonal expansion of Vgamma9-Vdelta2 (VgammaII family) subset indicating that, this expanded population of cells could be responsible for eliciting an immune response against oral tumor cells.