RÉSUMÉ
A 78-year-old woman with rheumatoid arthritis, who was started on baricitinib five or six months earlier, was referred to our hospital due to a subcutaneous abscess in her right axilla. Contrast-enhanced chest, abdomen, and pelvis computed tomography showed subcutaneous abscesses in her right axilla and lymphadenopathy with calcification. Cultures from the subcutaneous abscess and skin biopsy specimens were positive for Mycobacterium tuberculosis. These findings led to the diagnosis of scrofuloderma associated with tuberculous lymphadenitis. She was started on an antitubercular regimen of isoniazid, rifampicin, pyrazinamide, and ethambutol as the initial phase treatment (first 2 months), followed by isoniazid and rifampicin for 4 months (total 6 months). After 6 months of antitubercular treatment, the abscesses and lymphadenitis disappeared. Although cases of tuberculosis during JAK inhibitor treatment are rare, they are serious adverse events that require caution.
Sujet(s)
Antituberculeux , Polyarthrite rhumatoïde , Inhibiteurs des Janus kinases , Pyrazoles , Sulfonamides , Tuberculose ganglionnaire , Humains , Polyarthrite rhumatoïde/traitement médicamenteux , Polyarthrite rhumatoïde/complications , Femelle , Sujet âgé , Inhibiteurs des Janus kinases/effets indésirables , Antituberculeux/effets indésirables , Antituberculeux/administration et posologie , Sulfonamides/effets indésirables , Sulfonamides/administration et posologie , Pyrazoles/effets indésirables , Tuberculose ganglionnaire/traitement médicamenteux , Tuberculose ganglionnaire/diagnostic , Purines/effets indésirables , Purines/administration et posologie , Azétidines/effets indésirables , Azétidines/administration et posologie , Tuberculose cutanée/diagnostic , Tuberculose cutanée/traitement médicamenteux , Résultat thérapeutique , Mycobacterium tuberculosis/isolement et purification , Association de médicaments , Isoniazide/effets indésirables , Isoniazide/administration et posologieRÉSUMÉ
Pancreatic cancer is one of the leading causes of cancerrelated mortality and has the lowest 5year survival rate. Therefore, novel strategies are urgently required to treat pancreatic cancer. Pancreatic ductal adenocarcinoma (PDAC) cells rely on enhanced lysosomal function for survival and proliferation to facilitate the degradation of contents accumulated via autophagy and macropinocytosis. Previously, we have reported that the combination of epidermal growth factor receptor/HER2 inhibitor lapatinib and sphingosine analog fingolimod (FTY720) confers a significant cytostatic effect in lung cancer cells. In the present study, the combined effects of these drugs on PDAC cell lines, BxPC3, KP4, PANC1 and MIA PaCa2, were examined. It was observed that FTY720 enhanced the lapatinibinduced cytotoxic effect and caused noncanonical and lysosomedependent death in PDAC cells. Lapatinib and FTY720 induced lysosomal swelling and inhibited lysosomal acidification. Combination treatment with lapatinib and FTY720 increased lysosomal membrane permeability, induced mitochondrial depolarization, induced endoplasmic reticulum stress and disturbed intracellular calcium homeostasis. Additionally, the cytotoxic effect of lapatinib was enhanced by hydroxychloroquine or the CDK4/6 inhibitor abemaciclib, both of which induce lysosomal dysfunction. Collectively, these results indicated that the lysosometargeted drug combination induces multiple organelle dysfunction and exerts a marked cytotoxic effect in PDAC cells.
Sujet(s)
Carcinome du canal pancréatique/traitement médicamenteux , Chlorhydrate de fingolimod/pharmacologie , Lapatinib/pharmacologie , Lysosomes/effets des médicaments et des substances chimiques , Tumeurs du pancréas/traitement médicamenteux , Aminopyridines/pharmacologie , Antinéoplasiques/pharmacologie , Benzimidazoles/pharmacologie , Lignée cellulaire tumorale , Synergie des médicaments , Stress du réticulum endoplasmique/effets des médicaments et des substances chimiques , Humains , Hydroxychloroquine/pharmacologie , Modulateurs des récepteurs de la sphingosine 1 phosphate/pharmacologieRÉSUMÉ
Autophagy, a self-digestive system for cytoplasmic components, is required to maintain the amino acid pool for cellular homeostasis. We previously reported that the macrolide antibiotics azithromycin (AZM) and clarithromycin (CAM) have an inhibitory effect on autophagy flux, and they potently enhance the cytocidal effect of various anticancer reagents in vitro. This suggests that macrolide antibiotics can be used as an adjuvant for cancer chemotherapy. Since cancer cells require a larger metabolic demand than normal cells because of their exuberant growth, upregulated autophagy in tumor cells has now become the target for cancer therapy. In the present study, we examined whether macrolides exhibit cytotoxic effect under an amino acid-starving condition in head and neck squamous cancer cell lines such as CAL 27 and Detroit 562 as models of solid tumors with an upregulated autophagy in the central region owing to hypovascularity. AZM and CAM induced cell death under the amino acid-depleted (AAD) culture condition in these cell lines along with CHOP upregulation, although they showed no cytotoxicity under the complete culture medium. CHOP knockdown by siRNA in the CAL 27 cells significantly suppressed macrolide-induced cell death under the AAD culture condition. CHOP-/- murine embryonic fibroblast (MEF) cell lines also attenuated AZM-induced cell death compared with CHOP+/+ MEF cell lines. Using a tet-off atg5 MEF cell line, knockout of atg5, an essential gene for autophagy, also induced cell death and CHOP in the AAD culture medium but not in the complete culture medium. This suggest that macrolide-induced cell death via CHOP induction is dependent on autophagy inhibition. The cytotoxicity of macrolide with CHOP induction was completely cancelled by the addition of amino acids in the culture medium, indicating that the cytotoxicity is due to the insufficient amino acid pool. These data suggest the possibility of using macrolides for "tumor-starving therapy".
Sujet(s)
Antibactériens/pharmacologie , Autophagie/effets des médicaments et des substances chimiques , Azithromycine/pharmacologie , Carcinome épidermoïde/anatomopathologie , Mort cellulaire/effets des médicaments et des substances chimiques , Clarithromycine/pharmacologie , Tumeurs de la tête et du cou/anatomopathologie , Macrolides/pharmacologie , Animaux , Lignée cellulaire tumorale , Humains , SourisRÉSUMÉ
The ubiquitin-proteasome and autophagy-lysosome pathways are two major self-digestive systems for cellular proteins. Ubiquitinated misfolded proteins are degraded mostly by proteasome. However, when ubiquitinated proteins accumulate beyond the capacity of proteasome clearance, they are transported to the microtubule-organizing center (MTOC) along the microtubules to form aggresomes, and subsequently some of them are degraded by the autophagy-lysosome system. We previously reported that macrolide antibiotics such as azithromycin and clarithromycin block autophagy flux, and that concomitant treatment with the proteasome inhibitor bortezomib (BZ) and macrolide enhances endoplasmic reticulum (ER) stress-mediated apoptosis in breast cancer cells. As ubiquitinated proteins are concentrated at the aggresome upon proteasome failure, we focused on the microtubule as the scaffold of this transport pathway for aggresome formation. Treatment of metastatic breast cancer cell lines (e.g., MDA-MB231 cells) with BZ resulted in induction of aggresomes, which immunocytochemistry detected as a distinctive eyeball-shaped vimentin-positive inclusion body that formed in a perinuclear lesion, and that electron microscopy detected as a sphere of fibrous structure with some dense amorphous deposit. Vinorelbine (VNR), which inhibits microtubule polymerization, more effectively suppressed BZ-induced aggresome formation than paclitaxel (PTX), which stabilizes microtubules. Combined treatment using BZ and VNR, but not PTX, enhanced the cytotoxic effect and apoptosis induction along with pronounced ER stress loading such as upregulation of GRP78 and CHOP/GADD153. The addition of azithromycin to block autophagy flux in the BZ plus VNR-containing cell culture further enhanced the cytotoxicity. These data suggest that suppression of BZ-induced aggresome formation using an inhibitory drug such as VNR for microtubule polymerization is a novel strategy for metastatic breast cancer therapy.
Sujet(s)
Apoptose/effets des médicaments et des substances chimiques , Bortézomib/pharmacologie , Tumeurs du sein/anatomopathologie , Stress du réticulum endoplasmique/effets des médicaments et des substances chimiques , Proteasome endopeptidase complex/effets des médicaments et des substances chimiques , Vinblastine/analogues et dérivés , Antinéoplasiques/pharmacologie , Autophagie/effets des médicaments et des substances chimiques , Technique de Western , Tumeurs du sein/traitement médicamenteux , Tumeurs du sein/métabolisme , Prolifération cellulaire/effets des médicaments et des substances chimiques , Synergie des médicaments , Chaperonne BiP du réticulum endoplasmique , Femelle , Cytométrie en flux , Technique d'immunofluorescence , Humains , Cellules cancéreuses en culture , Vinblastine/pharmacologie , VinorelbineRÉSUMÉ
The inhibitory effects of macrolide antibiotics including clarithromycin (CAM) on autophagy flux have been reported. Although a macrolide antibiotic exhibits no cytotoxicity, its combination with bortezomib (BZ), a proteasome inhibitor, for the simultaneous blocking of the ubiquitin (Ub)proteasome and autophagylysosome pathways leads to enhanced multiple myeloma (MM) cell apoptosis induction via stress overloading of the endoplasmic reticulum (ER). As misfolded protein cargo is recruited by histone deacetylase 6 (HDAC6) to dynein motors for aggresome transport, serving to sequester misfolded proteins, we further investigated the cellular effects of targeting proteolytic pathways and aggresome formation concomitantly in MM cells. Pronounced apoptosis was induced by the combination of vorinostat [suberoylanilide hydroxamic acid (SAHA); potently inhibits HDAC6] with CAM and BZ compared with each reagent or a 2reagent combination. CAM/BZ treatment induced vimentin positiveaggresome formation along with the accumulation of autolysosomes in the perinuclear region, whereas they were inhibited in the presence of SAHA. The SAHA/CAM/BZ combination treatment maximally upregulated genes related to ER stress including C/EBP homologous protein (CHOP). Similarly to MM cell lines, enhanced cytotoxicity with CHOP upregulation following SAHA/CAM/BZ treatment was shown by a wildtype murine embryonic fibroblast (MEF) cell line; however, a CHOPdeficient MEF cell line almost completely canceled this pronounced cytotoxicity. Knockdown of HDAC6 with siRNA exhibited further enhanced CAM/BZinduced cytotoxicity and CHOP induction along with the cancellation of aggresome formation. Targeting the integrated networks of aggresome, proteasome, and autophagy is suggested to induce efficient ER stressmediated apoptosis in MM cells.
Sujet(s)
Apoptose/effets des médicaments et des substances chimiques , Autophagie/effets des médicaments et des substances chimiques , Stress du réticulum endoplasmique/effets des médicaments et des substances chimiques , Myélome multiple/traitement médicamenteux , Animaux , Protocoles de polychimiothérapie antinéoplasique , Acides boroniques/administration et posologie , Bortézomib , Lignée cellulaire tumorale , Clarithromycine/administration et posologie , Synergie des médicaments , Réticulum endoplasmique/effets des médicaments et des substances chimiques , Histone deacetylase 6 , Histone deacetylases/effets des médicaments et des substances chimiques , Humains , Acides hydroxamiques/administration et posologie , Souris , Myélome multiple/génétique , Myélome multiple/anatomopathologie , Proteasome endopeptidase complex/effets des médicaments et des substances chimiques , Pyrazines/administration et posologie , VorinostatRÉSUMÉ
MicroRNA (miR)-17-92a expression plays a crucial role in lymphocyte ontogeny. We therefore set out to determine miR-92a expression levels in peripheral blood lymphocytes from healthy subjects to ascertain any association between these levels and ageing. We found a positive correlation between the miR-92a expression level and the percentages of RO-CD8+CD27+ (P = 0.0046) and CD3+CD8+CD62L+ (P = 0.0011). This suggests that the majority of miR-92a of CD8+ T cells is derived from naïve cells, and the miR-92a expression level in CD8+ T cells declines progressively with age. These results indicate that the age-related attrition of naïve T cells is linked to a reduction of miR-92a in human T -lymphocytes. Therefore, we should careful attention when evaluating human miRNA levels in T lymphocytes to use normal control values.
RÉSUMÉ
Ultraviolet (UV) radiation is one of the most important environmental factors involved in the pathogenesis of premature skin ageing, termed photoageing. The harmful effects of UV in photoageing are associated with the generation of reactive oxygen species, and cellular antioxidants act to prevent the occurrence and reduce the severity of UV-induced photoageing. The transcription factor Nrf2 and its cytoplasmic anchor protein, Keap1, are central regulators of the cellular antioxidant response. Here, we investigated the role of the Nrf2-Keap1 pathway in photoageing using nrf2 gene-deficient (nrf2(-/-)) mice. Our results indicated that UVB-irradiated nrf2(-/-) mice showed accelerated photoageing, such as coarse wrinkle formation, loss of skin flexibility, epidermal thickening and deposition of extracellular matrix in the upper dermis. In addition, nrf2(-/-) mice also showed an increase in cutaneous reactivity for the lipid peroxidation product 4-hydroxy-2-nonenal and a significant decrease in cutaneous glutathione level. These findings indicate that Nrf2 plays the important role in the protection against UVB-induced photoageing.
Sujet(s)
Facteur-2 apparenté à NF-E2/déficit , Vieillissement de la peau/anatomopathologie , Vieillissement de la peau/effets des radiations , Peau/anatomopathologie , Peau/effets des radiations , Rayons ultraviolets/effets indésirables , Protéines adaptatrices de la transduction du signal/métabolisme , Aldéhydes/métabolisme , Animaux , Antioxydants/métabolisme , Protéines du cytosquelette/métabolisme , Femelle , Glutathion/métabolisme , Hypertrophie/anatomopathologie , Protéine-1 de type kelch associée à ECH , Peroxydation lipidique/effets des radiations , Souris , Souris de lignée ICR , Souris knockout , Souches mutantes de souris , Modèles animaux , Facteur-2 apparenté à NF-E2/génétique , Facteur-2 apparenté à NF-E2/métabolisme , Transduction du signal/physiologie , Peau/métabolisme , Facteurs tempsRÉSUMÉ
The serum concentration of thalidomide in multiple myeloma (MM) patients with renal insufficiency has not been investigated in Japan. We examined the serum concentration of thalidomide 12 and 16 hours after administration in 5 patients with MM (one with normal renal function and four with renal insufficiency, including one on hemodialysis (HD)) taking 100-200 mg/day. Concentrations 12 and 16 hours after administration corresponded to those before and after HD, respectively, in the patient on HD. The thalidomide concentration was not significantly increased by renal insufficiency. We could not detect any correlation between the concentration of thalidomide and its clinical effect. Since the elimination of thalidomide during HD seems to be greater than during the same period on a non-HD day, it was suggested that thalidomide could be eliminated by HD. The concentrations 12 hours after administration were similar with or without HD. From these results, even in Japanese patients, the thalidomide dosage need not be modified for renal insufficiency and HD.
Sujet(s)
Immunosuppresseurs/administration et posologie , Immunosuppresseurs/sang , Myélome multiple/traitement médicamenteux , Myélome multiple/métabolisme , Insuffisance rénale/complications , Insuffisance rénale/métabolisme , Thalidomide/administration et posologie , Thalidomide/sang , Sujet âgé , Asiatiques , Humains , Immunosuppresseurs/pharmacocinétique , Rein/métabolisme , Mâle , Adulte d'âge moyen , Myélome multiple/complications , Dialyse rénale , Thalidomide/pharmacocinétiqueRÉSUMÉ
Ultraviolet (UV) radiation is one of the most important environmental factors involved in the pathogenesis of skin aging and cancer. Many harmful effects of UV radiation are associated with the generation of reactive oxygen species, and cellular antioxidants act to prevent the occurrence and reduce the severity of UV-induced skin disorders. Transcription factor NF-E2-related Factor 2 (Nrf2) and its cytoplasmic anchor protein Kelch-like-ECH-associated protein 1 (Keap1) are central regulators of the cellular antioxidant response. In this study, we investigated the effects of UV irradiation on the activation of Nrf2 in dermal fibroblasts. We found that UVA irradiation, but not UVB, causes nuclear translocation and accumulation of Nrf2 by a factor of 6.5 as compared with unirradiated controls. The nuclear accumulation of Nrf2 induced by UVA was enhanced by the photosensitizer hematoporphyrin. To evaluate the protective role of Nrf2 against UVA radiation, we examined UVA-induced apoptosis using dermal fibroblasts derived from nrf2 or keap1 gene knockout mice. Whereas disruption of nrf2 increased the number of apoptotic cells following UVA irradiation by 1.7-fold, disruption of keap1 decreased the apoptotic cell number by half as compared with wild-type controls. These findings thus demonstrate that the Nrf2-Keap1 pathway plays an important role in the protection of the skin against UVA irradiation.
Sujet(s)
Apoptose/effets des médicaments et des substances chimiques , Protéines de liaison à l'ADN/métabolisme , Derme/effets des radiations , Fibroblastes/effets des radiations , Transactivateurs/métabolisme , Rayons ultraviolets/effets indésirables , Protéines adaptatrices de la transduction du signal/génétique , Animaux , Noyau de la cellule/métabolisme , Cellules cultivées , Protéines du cytosquelette/génétique , Derme/anatomopathologie , Derme/physiologie , Fibroblastes/anatomopathologie , Fibroblastes/physiologie , Régulation de l'expression des gènes codant pour des enzymes/effets des radiations , Glutamate-cysteine ligase/génétique , Hématoporphyrines/pharmacologie , Protéine-1 de type kelch associée à ECH , Souris , Souris de lignée ICR , Souches mutantes de souris , Facteur-2 apparenté à NF-E2RÉSUMÉ
Loricrin is a major component of the epidermal cornified cell envelope, and is expressed only in terminally differentiated keratinocytes. This cell differentiation-specific expression pattern suggests specific suppression of loricrin gene expression in undifferentiated keratinocytes as well as its activation in differentiated keratinocytes. We identified a negative regulatory sequence element in the first intron of the mouse loricrin gene involved in suppression of loricrin gene expression in undifferentiated keratinocytes. A database search indicated that this sequence contained the putative inverted Yin-Yang 1 (YY1)-binding motif. Constructs with point mutations in the putative YY1-binding motif showed increased reporter activity, indicating that YY1 negatively regulates loricrin gene transcription. Co-transfection experiments using a YY1 expression vector revealed that YY1 represses loricrin promoter activity. Western blotting and immunohistochemical analyses indicated that YY1 is more abundant in undifferentiated than in differentiated keratinocytes. These findings suggest that YY1 contributes to specific loricrin gene expression in differentiated keratinocytes by suppression of its transcription in undifferentiated keratinocytes. Furthermore, we demonstrated that forced expression of YY1 in differentiated keratinocytes results in specific downregulation of expression of other early and late differentiation markers.