Identification of an IL-1 receptor mutation driving autoinflammation directs IL-1-targeted drug design.

The interleukin 1 (IL-1) pathway signals through IL-1 receptor type 1 (IL-1R1) and emerges as a central mediator for systemic inflammation. Aberrant IL-1 signaling leads to a range of autoinflammatory diseases. Here, we identified a de novo missense variant in IL-1R1 (p.Lys131Glu) in a patient with chronic recurrent multifocal osteomyelitis (CRMO). Patient PBMCs showed strong inflammatory signatures, particularly in monocytes and neutrophils. The p.Lys131Glu substitution affected a critical positively charged amino acid, which disrupted the binding of the antagonist ligand, IL-1Ra, but not IL-1α or IL-1ß. This resulted in unopposed IL-1 signaling. Mice with a homologous mutation exhibited similar hyperinflammation and greater susceptibility to collagen antibody-induced arthritis, accompanied with pathological osteoclastogenesis. Leveraging the biology of the mutation, we designed an IL-1 therapeutic, which traps IL-1ß and IL-1α, but not IL-1Ra. Collectively, this work provides molecular insights and a potential drug for improved potency and specificity in treating IL-1-driven diseases.

A gain-of-function variation in PLCG1 causes a new immune dysregulation disease.

BACKGROUND: Phospholipase C (PLC) γ1 is a critical enzyme regulating nuclear factor-κB (NF-κB), extracellular signal-related kinase, mitogen-activated protein kinase, and nuclear factor of activated T cells signaling pathways, yet germline PLCG1 mutation in human disease has not been reported. OBJECTIVE: We aimed to investigate the molecular pathogenesis of a PLCG1 activating variant in a patient with immune dysregulation. METHODS: Whole exome sequencing was used to identify the patient's pathogenic variants. Bulk RNA sequencing, single-cell RNA sequencing, quantitative PCR, cytometry by time of flight, immunoblotting, flow cytometry, luciferase assay, IP-One ELISA, calcium flux assay, and cytokine measurements in patient PBMCs and T cells and COS-7 and Jurkat cell lines were used to define inflammatory signatures and assess the impact of the PLCG1 variant on protein function and immune signaling. RESULTS: We identified a novel and de novo heterozygous PLCG1 variant, p.S1021F, in a patient presenting with early-onset immune dysregulation disease. We demonstrated that the S1021F variant is a gain-of-function variant, leading to increased inositol-1,4,5-trisphosphate production, intracellular Ca2+ release, and increased phosphorylation of extracellular signal-related kinase, p65, and p38. The transcriptome and protein expression at the single-cell level revealed exacerbated inflammatory responses in the patient's T cells and monocytes. The PLCG1 activating variant resulted in enhanced NF-κB and type II interferon pathways in T cells, and hyperactivated NF-κB and type I interferon pathways in monocytes. Treatment with either PLCγ1 inhibitor or Janus kinase inhibitor reversed the upregulated gene expression profile in vitro. CONCLUSIONS: Our study highlights the critical role of PLCγ1 in maintaining immune homeostasis. We illustrate immune dysregulation as a consequence of PLCγ1 activation and provide insight into therapeutic targeting of PLCγ1.

Integration of Topological Insulator Josephson Junctions in Superconducting Qubit Circuits.

The integration of semiconductor Josephson junctions (JJs) in superconducting quantum circuits provides a versatile platform for hybrid qubits and offers a powerful way to probe exotic quasiparticle excitations. Recent proposals for using circuit quantum electrodynamics (cQED) to detect topological superconductivity motivate the integration of novel topological materials in such circuits. Here, we report on the realization of superconducting transmon qubits implemented with (Bi0.06Sb0.94)2Te3 topological insulator (TI) JJs using ultrahigh vacuum fabrication techniques. Microwave losses on our substrates, which host monolithically integrated hardmasks used for the selective area growth of TI nanostructures, imply microsecond limits to relaxation times and, thus, their compatibility with strong-coupling cQED. We use the cavity-qubit interaction to show that the Josephson energy of TI-based transmons scales with their JJ dimensions and demonstrate qubit control as well as temporal quantum coherence. Our results pave the way for advanced investigations of topological materials in both novel Josephson and topological qubits.

Role of Circular RNAs in the Pathogenesis of Malignant Melanoma.

Malignant melanoma is one of the most aggressive types of skin cancer. Thus, efficient diagnosis and treatment methods are crucial for advanced melanoma. Circular RNAs (circRNAs) have been regarded as a 'splicing noise' in the past decades. However, several circRNAs have been recently reported to be differentially expressed in melanoma, and the cell or tissue-specific expression makes these suitable candidate diagnostic or therapeutic biomarkers. In addition, emerging studies have confirmed that circular RNAs play pivotal roles in the proliferation, invasion, metastasis, and migration of malignant melanoma. However, specific pathogenic mechanisms between melanoma and circRNAs remain unclear. In the present study, it was summarized that circRNAs are associated with the pathogenesis of melanoma, including hsa_circ_0083444, hsa_circ_0005320, hsa_circ_0067531, hsa_circ_0084043, hsa_circ_0000082, hsa_circ_0016418, hsa_circ_0085533 and hsa_circ_0025039, hsa_circ_0001946, hsa_circ_0002770, hsa_circ_0079593, hsa_circ_0027247, hsa_circ_0017247, hsa_circ_0020710. These can provide potential diagnosis, treatment, and prognostication biomarkers for advanced melanoma in clinical applications.

Glutamine deficiency promotes recurrence and metastasis in colorectal cancer through enhancing epithelial-mesenchymal transition.

BACKGROUND: Glutamine is the most abundant amino acid in the body and plays a vital role in colorectal cancer (CRC) cell metabolism. However, limited studies have investigated the clinical and prognostic significance of preoperative serum glutamine levels in patients with colorectal cancer, and the underlying mechanism has not been explored. METHODS: A total of 121 newly diagnosed CRC patients between 2012 and 2016 were enrolled in this study. Serum glutamine levels were detected, and their associations with clinicopathological characteristics, systemic inflammation markers, carcinoembryonic antigen (CEA) and prognosis were analysed. In addition, the effect of glutamine depletion on recurrence and metastasis was examined in SW480 and DLD1 human CRC cell lines, and epithelial-mesenchymal transition (EMT)-related markers were detected to reveal the possible mechanism. RESULTS: A decreased preoperative serum level of glutamine was associated with a higher T-class and lymph node metastasis (P < 0.05). A higher serum level of glutamine correlated with a lower CEA level (r = - 0.25, P = 0.02). Low glutamine levels were correlated with shorter overall survival (OS) and disease-free survival (DFS). Multivariate Cox regression analysis showed that serum glutamine was an independent prognostic factor for DFS (P = 0.018), and a nomogram predicting the probability of 1-, 3- and 5-year DFS after radical surgery was built. In addition, glutamine deficiency promoted the migration and invasion of CRC cells. E-cadherin, a vital marker of EMT, was decreased, and EMT transcription factors, including zeb1and zeb2, were upregulated in this process. CONCLUSIONS: This study elucidated that preoperative serum glutamine is an independent prognostic biomarker to predict CRC progression and suggested that glutamine deprivation might promote migration and invasion in CRC cells by inducing the EMT process.

Identification of oral precancerous and cancerous tissue by swept source optical coherence tomography.

BACKGROUND AND OBJECTIVES: Distinguishing cancer from precancerous lesions is critical and challenging in oral medicine. As a noninvasive method, optical coherence tomography (OCT) has the advantages of real-time, in vivo, and large-depth imaging. Texture information hidden in OCT images can provide an important auxiliary effect for improving diagnostic accuracy. The aim of this study is to explore a reliable and accurate OCT-based method for the screening and diagnosis of human oral diseases, especially oral cancer. MATERIALS AND METHODS: Fresh ex vivo oral tissues including normal mucosa, leukoplakia with epithelial hyperplasia (LEH), and oral squamous cell carcinoma (OSCC) were imaged intraoperatively by a homemade OCT system, and 58 texture features were extracted to create computational models of these tissues. A principal component analysis algorithm was employed to optimize the combination of texture feature vectors. The identification based on artificial neural network (ANN) was proposed and the sensitivity/specificity was calculated statistically to evaluate the classification performance. RESULTS: A total of 71 sites of three types of oral tissues were measured, and 5176 OCT images of three types of oral tissues were used in this study. The superior classification result based on ANN was obtained with an average accuracy of 98.17%. The sensitivity and specificity of normal mucosa, LEH, and OSCC are 98.17% / 98.38%, 93.81% / 98.54%, and 98.11% / 99.04%, respectively. CONCLUSION: It is demonstrated from the high accuracies, sensitivities, and specificities that texture-based analysis can be used to identify oral precancerous and cancerous tissue in OCT images, and it has the potential to help surgeons in diseases screening and diagnosis effectively.

Classification of oral salivary gland tumors based on texture features in optical coherence tomography images.

Currently, the diagnoses of oral diseases primarily depend on the visual recognition of experienced clinicians. It has been proven that automatic recognition based on images can support clinical decision-making by extracting and analyzing objective hidden information. In recent years, optical coherence tomography (OCT) has become a powerful optical imaging technique with the advantages of high resolution and non-invasion. In our study, a dataset composed of four kinds of oral salivary gland tumors (SGTs) was obtained from a homemade swept-source OCT, including two benign and two malignant tumors. Seventy-six texture features were extracted from OCT images to create computational models of diseases. It was demonstrated that the artificial neural network (ANN) based on principal component analysis (PCA) can obtain high diagnostic sensitivity and specificity (higher than 99%) for these four kinds of tumors. The classification accuracy of each tumor is larger than 99%. In addition, the performances of two classifiers (ANN and support vector machine) were quantitatively evaluated based on SGTs. It was proven that the texture features in OCT images provided objective information to classify oral tumors.

Hyaluronan injection versus oral glucosamine and diclofenac in the treatment of temporomandibular joint osteoarthritis.

OBJECTIVES: This study was aimed to compare the effects of 4 biweekly hyaluronan (HA) injection with glucosamine and diclofenac oral administration on TMJ OA patients. MATERIALS AND METHODS: This retrospective cohort study included TMJ OA patients who had the treatment of 4 biweekly HA injection (group HA) or oral glucosamine hydrochloride for 3 months and diclofenac sodium for 2 weeks (group G/D), and had complete data at first-visit, 3 months, 6 months, and 12 months. Clinical signs and symptoms were scored by anamnestic dysfunction index (Ai) and clinical dysfunction index (Di), and condylar bone changes were evaluated by CBCT scoring system. RESULTS: We included 22 patients in group HA and 20 patients in group G/D. After HA injection, Ai was decreased from 4.3 to 1.6(CI [- 4.0, - 1.4]) at 3-month follow-up, which was smaller than that in group G/D significantly. Di in group HA was declined significantly from 8.1 at first-visit to 3.6 at 3-month follow-up, while Di in group G/D scarcely changed until at 6- and 12-month follow-up. Neither HA injection nor oral glucosamine/diclofenac showed positive effect on the bone of TMJs during follow-ups with statistical significance. CONCLUSIONS: HA injection alleviated signs and symptoms of TMJ OA rapidly and presented superior clinical effects over oral glucosamine with diclofenac. However, both treatments did not limit the bone destruction of TMJs significantly. CLINICAL RELEVANCE: This cohort study provides knowledge on the symptom relief and bone changes of TMJ OA patients when treated with HA injection or glucosamine and diclofenac oral administration.

Application of biochar activated persulfate in the treatment of typical azo pigment wastewater.

With the increase of the azo pigment wastewater, it is necessary to seek an efficient and sustainable treatment method to address issues of damaging water ecosystems and human health. In this work, organic representing azo dye Acid Orange 7 (AO7), heavy metal representing hexavalent chromium (Cr(VI)), and inorganic representing ammonia nitrogen (NH4+-N) were selected to roughly simulate the azo pigment wastewater. The simultaneous decontamination of multi-target pollutants by 700 °C pyrolyzed peanut shell biochar (BC) with persulfate (PDS) was evaluated. The results showed that AO7, Cr(VI) and NH4+-N could finally reach 100%, 85% and 30% removal ratios separately in the BC/PDS/mixed pollutants system under certain basic conditions. Functional groups (hydroxyl groups (C-OH) and carboxylic ester/lactone groups (O-C=O)) were found by XPS as competing sites for adsorption and activation and were gradually consumed as the reaction proceeded. Combining a series of experiments results and EPR analysis, it was found that AO7 removal worked best and it relied on both the radical pathway (including SO4•-, •OH, O2-•, but not 1O2) and adsorption. Cr(VI) was mainly adsorbed and reduced by BC surface to form Cr(OH)3 and Cr2O3, and the remaining part could be reduced by O2-•, followed by •OH. NH4+-N was removed primarily by the radical same as AO7. Meanwhile, the three target pollutants have a co-competitive mechanism. Specifically, they competed for radicals and adsorption sites simultaneously, while the presence of AO7 and NH4+-N would consume the generated oxidizing radicals and further promote the removal of Cr(VI). The fixed-bed reactor simulated the continuous treatment of wastewater. Various anions (chloride (Cl-), nitrate (NO3-), carbonate (CO32-), and hydrogen phosphate (HPO42-)) interfered differently with the pollutant removal. These findings demonstrate a new dimension of BC potential for decontamination of azo pigment wastewater.

Classification of Salivary Gland Tumors Based on Quantitative Optical Coherence Tomography.

BACKGROUND AND OBJECTIVES: Visual inspection is the primary diagnostic method for oral diseases, and its accuracy of diagnosis mainly depends on surgeons' experience. Histological examination is still the golden standard, but it is invasive and time-consuming. In order to address these issues, as a noninvasive imaging technique, optical coherence tomography (OCT) can differentiate oral tissue with advantages of real-time, in situ, and high resolution. The aim of this study is to explore optimal quantitative parameters in OCT images to distinguish different salivary gland tumors. STUDY DESIGN/MATERIALS AND METHODS: OCT images of four salivary gland tumors were obtained from 14 patients, including mucoepidermoid carcinoma (MC), adenoid cystic carcinoma (ACC), basal cell adenoma (BCA), and pleomorphic adenoma (PA). Two parameters of optical attenuation coefficient (OAC) and standard deviation (SD) along the depth of OCT signal were combined to create a computational model of classification, and sensitivity/specificity of classification was calculated statistically to evaluate their results. RESULTS: A total of 5,919 two-dimensional (2D) OCT images were used for quantitative analysis. The classification sensitivities of 89.6%, 95.0%, 89.5%, 97.8%, and specificities of 97.6%, 99.0%, 98.0%, 98.2%, respectively, were obtained for MC, ACC, BCA, and PA, with the thresholds of 3.6 mm-1 based on OAC and 0.22/0.18 based on SD. CONCLUSION: It was demonstrated that OAC and SD could be considered as important parameters in quantitative analysis of OCT images for salivary gland tissue characterization and intraoperative diagnosis. It is of great potential value in promoting the application of this method based on OCT in clinical practice. Lasers Surg. © 2020 Wiley Periodicals LLC.

A meta-analysis of fractional CO2 laser combined with PRP in the treatment of acne scar.

This study aimed to analyze the effectiveness and safety of ablative fractional carbon dioxide laser systems (CO2 AFL) combined with autologous platelet-rich plasma (PRP) in the treatment of acne scars through the retrieval and collection of related literature to further guide the treatment of acne scars. We searched Web of Science, PubMed, Embase, Wanfang Data, Chinese National Knowledge Infrastructure, and VIP Database. All randomized and nonrandomized controlled trials on CO2 AFL combined with PRP in the treatment of acne scars were included, and Revman5.3 systematic review software was used in the meta-analysis. Nine studies were included in this meta-analysis. The data analysis results showed that the CO2 AFL combined with PRP treatment group showed significantly better results than the pure CO2 AFL control group in terms of clinical improvement score, clinical improvement rate, patient satisfaction, and crusting period. The results of this meta-analysis showed that CO2 AFL combined with PRP in the treatment of acne scars is more effective and safer than CO2 AFL alone.

Activation of the ABA Signal Pathway Mediated by GABA Improves the Drought Resistance of Apple Seedlings.

Drought seriously affects the yield and quality of apples. γ-aminobutyric acid (GABA) plays an important role in the responses of plants to various stresses. However, the role and possible mechanism of GABA in the drought response of apple seedlings remain unknown. To explore the effect of GABA on apple seedlings under drought stress, seedlings of Malus hupehensis were treated with seven concentrations of GABA, and the response of seedlings under 15-day drought stress was observed. The results showed that 0.5 mM GABA was the most effective at relieving drought stress. Treatment with GABA reduced the relative electrical conductivity and MDA content of leaves induced by drought stress and significantly increased the relative water content of leaves. Exogenous GABA significantly decreased the stomatal conductance and intercellular carbon dioxide concentration and transpiration rate, and it significantly increased the photosynthetic rate under drought. GABA also reduced the accumulation of superoxide anions and hydrogen peroxide in leaf tissues under drought and increased the activities of POD, SOD, and CAT and the content of GABA. Exogenous treatment with GABA acted through the accumulation of abscisic acid (ABA) in the leaves to significantly decrease stomatal conductance and increase the stomatal closure rate, and the levels of expression of ABA-related genes PYL4, ABI1, ABI2, HAB1, ABF3, and OST1 changed in response to drought. Taken together, exogenous GABA can enhance the drought tolerance of apple seedlings.

High-Temperature Requirement A1 Protease as a Rate-Limiting Factor in the Development of Osteoarthritis.

Preserving the mature articular cartilage of joints is a critical focus in the prevention and treatment of osteoarthritis. We determined whether the genetic inactivation of high-temperature requirement A1 (HtrA1) can significantly attenuate the degradation of articular or condylar cartilage. Two types of mouse models of osteoarthritis were used, a spontaneous mutant mouse model [type XI collagen-haploinsufficient (Col11a1+/-) mice] and two post-traumatic mouse models [destabilization of the medial meniscus (DMM) on the knee and a partial discectomy (PDE) on the temporomandibular joint]. Three different groups of mice were generated: i) HtrA1 was genetically deleted from Col11a1+/- mice (HtrA1-/-;Col11a1+/-), ii) HtrA1-deficient mice (HtrA1-/-) were subjected to DMM, and iii) HtrA1-/- mice were subjected to PDE. Knee and temporomandibular joints from the mice were characterized for evidence of cartilage degeneration. The degradation of articular or condylar cartilage was significantly delayed in HtrA1-/-;Col11a1+/- mice and HtrA1-/- mice after DMM or PDE. The amount of collagen type VI was significantly higher in the articular cartilage in HtrA1-/-;Col11a1+/- mice, compared with that in Col11a1+/- mice. The genetic removal of HtrA1 may delay the degradation of articular or condylar cartilage in mice.

Using CT texture analysis to differentiate between peripheral lung cancer and pulmonary inflammatory pseudotumor.

BACKGROUND: This study is to distinguish peripheral lung cancer and pulmonary inflammatory pseudotumor using CT-radiomics features extracted from PET/CT images. METHODS: In this study, the standard 18F-fluorodeoxyglucose positron emission tomography/ computed tomography (18 F-FDG PET/CT) images of 21 patients with pulmonary inflammatory pseudotumor (PIPT) and 21 patients with peripheral lung cancer were retrospectively collected. The dataset was used to extract CT-radiomics features from regions of interest (ROI), The intra-class correlation coefficient (ICC) was used to screen the robust feature from all the radiomic features. Using, then, statistical methods to screen CT-radiomics features, which could distinguish peripheral lung cancer and PIPT. And the ability of radiomics features distinguished peripheral lung cancer and PIPT was estimated by receiver operating characteristic (ROC) curve and compared by the Delong test. RESULTS: A total of 435 radiomics features were extracted, of which 361 features showed relatively good repeatability (ICC ≥ 0.6). 20 features showed the ability to distinguish peripheral lung cancer from PIPT. these features were seen in 14 of 330 Gray-Level Co-occurrence Matrix features, 1 of 49 Intensity Histogram features, 5 of 18 Shape features. The area under the curves (AUC) of these features were 0.731 ± 0.075, 0.717, 0.748 ± 0.038, respectively. The P values of statistical differences among ROC were 0.0499 (F9, F20), 0.0472 (F10, F11) and 0.0145 (F11, Mean4). The discrimination ability of forming new features (Parent Features) after averaging the features extracted at different angles and distances was moderate compared to the previous features (Child features). CONCLUSION: Radiomics features extracted from non-contrast CT based on PET/CT images can help distinguish peripheral lung cancer and PIPT.

Identification of oral cancer in OCT images based on an optical attenuation model.

Surgery is still the first choice to treat oral cancer, where it is important to detect surgical margins in order to reduce cancer recurrence and maintain oral-maxillofacial function simultaneously. As a non-invasive and in situ imaging technique, optical coherence tomography (OCT) can obtain images close to the resolution of histopathology, which makes it have great potential in intraoperative diagnosis. However, it is not enough to find surgical margins accurately just observing OCT images directly and qualitatively. The purpose of this study is to identify oral cancer in OCT images by investigating the quantitative difference of cancer and non-cancer tissue. Based on an available optical attenuation model and the axial confocal PSF of a home-made swept source OCT system, by using fresh ex vivo human oral tissues from 14 patients of oral squamous cell carcinoma (OSCC) as the samples, diagnosis with sensitivity (97.88%) and specificity (83.77%) was achieved at the attenuation threshold of 4.7 mm-1, and the accuracy of identification reached 91.15% in our study. Our preliminary results demonstrated that the oral cancer resection will be guided accurately and the clinical application of OCT will be further promoted by deeply mining the information hidden in OCT images.

Overexpression of MdARD4 Accelerates Fruit Ripening and Increases Cold Hardiness in Tomato.

Ethylene plays an important role in stress adaptation and fruit ripening. Acireductone dioxygenase (ARD) is pivotal for ethylene biosynthesis. However, the response of ARD to fruit ripening or cold stress is still unclear. In this study, we identified three members of Malus ARD family, and expression profile analysis revealed that the transcript level of MdARD4 was induced during apple fruit ripening and after apple plants were being treated with cold stress. To investigate its function in cold tolerance and fruit ripening, MdARD4 was ectopically expressed in Solanum lycopersicum cultivar 'Micro-Tom', which has been considered as an excellent model plant for the study of fruit ripening. At the cellular level, the MdARD protein expressed throughout Nicotiana benthamiana epidermal cells. Overexpression of MdARD4 in tomato demonstrated that MdARD4 regulates the ethylene and carotenoid signaling pathway, increases ethylene and carotenoid concentrations, and accelerates fruit ripening. Furthermore, MdARD4 increased the antioxidative ability and cold hardiness in tomato. To conclude, MdARD4 may potentially be used in apple breeding to accelerate fruit ripening and increase cold hardiness.

Nickel-Catalyzed Decarboxylative Generation and Asymmetric Allylation of 2-Azaallyl Anions.

The first nickel-catalyzed asymmetric decarboxylative allylation (DcA) of allyl 2,2-diarylglycinate imines is reported. This transformation utilizes a chiral ferrocenyl bidentate ligand and a Ni(0) precatalyst to mediate the decarboxylative generation and asymmetric allylation of 2-azaallyl anions, affording α-aryl homoallylic imines in modest-to-high yields and moderate-to-high enantiomeric ratios. The resulting Ni-catalyzed transformation proved to be less general in comparison to our previously reported analogous Pd-mediated protocol, but it still exhibited certain advantages in regard to the regio- and enantioselectivity of the C-C bond formation.

Contrast analysis of the relationship between the HRCT sign and new pathologic classification in small ground glass nodule-like lung adenocarcinoma.

PURPOSE: To perform contrast analysis of the relationship between high-resolution computed tomography (HRCT) signs and new pathologic classification of small GGNs-like lung adenocarcinoma. MATERIALS AND METHODS: The HRCT data from 145 pathologically confirmed cases of small GGNs of lung adenocarcinoma were analysed retrospectively. The 145 cases of GGNs were divided into pre-invasive (PI) group (n = 46), micro-invasive adenocarcinoma (MIA) group (n = 48), and invasive adenocarcinoma (IAC) group (n = 51). HRCT imaging sign of GGNs in each group was assessed and compared. RESULTS: Significant differences in GGN size were found among the three groups (P < 0.05). The presence of a tumour-lung interface in the MIA and IAC groups was significantly higher than that in the PI group (P < 0.05), but no significant difference was found between the MIA and IAC groups. The presence of a pleural indentation sign in the IAC group was significantly higher than that in the other two groups (P < 0.05), but no significant difference was noted between the latter two groups. Significant differences were found in the lobulated and spicule signs among the three groups (P < 0.05). The presence of a microvascular sign in the MIA and IAC groups was significantly higher than that in the PI group (P < 0.05). No significant difference was found in the GGN density, vacuole sign, air bronchus sign and notch sign among the three groups. CONCLUSIONS: The HRCT signs of GGNs could be used to differentiate among pre-invasive lesions, micro-invasive lesions and invasive lung adenocarcinoma.

Cyclophosphamide enhances anti-tumor effects of a fibroblast activation protein α-based DNA vaccine in tumor-bearing mice with murine breast carcinoma.

Cyclophosphamide (CY) is a DNA alkylating agent, which is widely used with other chemotherapy drugs in the treatment of various types of cancer. It can be used not only as a chemotherapeutic but also as an immunomodulatory agent to inhibit IL-10 expression and T regulatory cells (Tregs). Fibroblast activation protein α (FAPα) is expressed in cancer-associated fibroblasts in the tumor microenvironment. Immunotherapy based on FAPα, as a tumor stromal antigen, typically induces specific immune response targeting the tumor microenvironment. This study evaluated the efficacy of a previously unreported CY combination strategy to enhance the limited anti-tumor effect of a DNA vaccine targeting FAPα. The results suggested CY administration could promote the percentage of splenic CD8+ T cells and decrease the proportion of CD4 + CD25 + Foxp3+ Tregs in spleen. In tumor tissues, levels of immunosuppressive cytokines including IL-10 and CXCL-12 were also reduced. Meanwhile, the CY combination did not impair the FAPα-specific immunity induced by the DNA vaccine and further reduced tumor stromal factors. Most importantly, FAP-vaccinated mice also treated with CY chemotherapy showed a marked suppression of tumor growth (inhibition ratio =80%) and a prolongation of survival time. Thus, the combination of FAPα immunotherapy and chemotherapy with CY offers new insights into improving cancer therapies.

Anti-tumor effects of DNA vaccine targeting human fibroblast activation protein α by producing specific immune responses and altering tumor microenvironment in the 4T1 murine breast cancer model.

Fibroblast activation protein α (FAPα) is a tumor stromal antigen overexpressed by cancer-associated fibroblasts (CAFs). CAFs are genetically more stable compared with the tumor cells and immunosuppressive components of the tumor microenvironment, rendering them excellent targets for cancer immunotherapy. DNA vaccines are widely applied due to their safety. To specifically destroy CAFs, we constructed and examined the immunogenicity and anti-tumor immune mechanism of a DNA vaccine expressing human FAPα. This vaccine successfully reduced 4T1 tumor growth through producing FAPα-specific cytotoxic T lymphocyte responses which could kill CAFs, and the decrease in FAPα-expressing CAFs resulted in markedly attenuated expression of collagen I and other stromal factors that benefit the tumor progression. Based on these results, a DNA vaccine targeting human FAPα may be an attractive and effective cancer immunotherapy strategy.