Lipid Nanoparticle-Associated Inflammation is Triggered by Sensing of Endosomal Damage: Engineering Endosomal Escape Without Side Effects.

Lipid nanoparticles (LNPs) have emerged as the dominant platform for RNA delivery, based on their success in the COVID-19 vaccines and late-stage clinical studies in other indications. However, we and others have shown that LNPs induce severe inflammation, and massively aggravate pre-existing inflammation. Here, using structure-function screening of lipids and analyses of signaling pathways, we elucidate the mechanisms of LNP-associated inflammation and demonstrate solutions. We show that LNPs' hallmark feature, endosomal escape, which is necessary for RNA expression, also directly triggers inflammation by causing endosomal membrane damage. Large, irreparable, endosomal holes are recognized by cytosolic proteins called galectins, which bind to sugars on the inner endosomal membrane and then regulate downstream inflammation. We find that inhibition of galectins abrogates LNP-associated inflammation, both in vitro and in vivo . We show that rapidly biodegradable ionizable lipids can preferentially create endosomal holes that are smaller in size and reparable by the endosomal sorting complex required for transport (ESCRT) pathway. Ionizable lipids producing such ESCRT-recruiting endosomal holes can produce high expression from cargo mRNA with minimal inflammation. Finally, we show that both routes to non-inflammatory LNPs, either galectin inhibition or ESCRT-recruiting ionizable lipids, are compatible with therapeutic mRNAs that ameliorate inflammation in disease models. LNPs without galectin inhibition or biodegradable ionizable lipids lead to severe exacerbation of inflammation in these models. In summary, endosomal escape induces endosomal membrane damage that can lead to inflammation. However, the inflammation can be controlled by inhibiting galectins (large hole detectors) or by using biodegradable lipids, which create smaller holes that are reparable by the ESCRT pathway. These strategies should lead to generally safer LNPs that can be used to treat inflammatory diseases.

Features of osteoporosis in male patients with bronchiectasis, a cross-sectional study.

Background: Osteoporosis increases the burden and disease related adverse events of comorbidities in some chronic disease. The relationships between osteoporosis and bronchiectasis are not fully understood. This cross-sectional study explores the features of osteoporosis in male patients with bronchiectasis. Methods: From January 2017 to December 2019, male patients (age >50 years) with stable bronchiectasis were included, as were normal subjects. Data on demographic characteristics and clinical features were collected. Results: Totally, 108 male patients with bronchiectasis and 56 controls were analyzed. Osteoporosis was observed in 31.5% (34/108) of patients with bronchiectasis and 17.9% (10/56) of controls (P=0.001). The T-score negatively correlated with age (R=-0.235, P=0.014) and bronchiectasis severity index score (BSI; R=-0.336, P<0.001). BSI score ≥9 was a major factor associated with osteoporosis [odd ratio (OR) =4.52; 95% confidence interval (CI): 1.57-12.96; P=0.005]. Other factors associated with osteoporosis included body-mass index (BMI) <18.5 kg/m2 (OR =3.44; 95% CI: 1.13-10.46; P=0.030), age ≥65 years (OR =2.87; 95% CI: 1.01-7.55; P=0.033), and a smoking history (OR =2.78; 95% CI: 1.04-7.47; P=0.042). Conclusions: The prevalence of osteoporosis was higher in male bronchiectasis patients than that in controls. Factors including age, BMI, smoking history, and BSI were associated with osteoporosis. Early diagnosis and treatment might be of great value in prevention and management of osteoporosis in patients with bronchiectasis.

The global, regional, and national burden of nasopharyngeal carcinoma and its attributable risk factors in 204 countries and territories, 1990-2019.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is uncommon in most areas of the world but poses a significant public health burden in endemic regions. OBJECTIVES: We provide an overview of the most recent global epidemiology of nasopharyngeal cancer (NPC). METHODS: We estimated the burden of NPC in 204 countries and territories by age, sex, and Socio-Demographic Index (SDI) from 1990 to 2019. RESULTS: At the GBD regional level, the most severe age-standardized incidence in 2019 occurred in East Asia. From 1990 to 2019, the East Asia and High-income Asia Pacific had the greatest increase in percentage in age-standardized incidence. Central Asia and the Caribbean had the greatest increase in percentage in age-standardized disability-adjusted life-years (DALY) and death rates. At the national level, Cabo Verde, Romania, and the Cyprus reported the largest percentage increases in the age-standardized incidence. Cabo Verde, Romania, and Jamaica reported the largest increases in the age-standardized DALY and death rates. CONCLUSIONS: The global age-standardized incidence of NPC increased globally between 1990 and 2019, especially in the East Asia.

Metformin attenuates cadmium-induced degeneration of spiral ganglion neuron via restoring autophagic flux in primary culture.

Cadmium (Cd), a common environmental and occupational toxicant, is an important risk factor for hearing loss. After exposure, Cd accumulates in the inner ear and induces spiral ganglion neuron (SGN) degeneration; however, the underlying mechanisms are poorly understood. Dysfunctional autophagy has been implicated in many neurodegenerative diseases, including Cd-induced neurotoxicity. Metformin has been validated to confer not only anti-hyperglycaemic but also neuroprotective effects. However, the relationship between autophagy dysfunction, SGN degeneration, and the effect of metformin on Cd-induced SGN neurotoxicity has not yet been established. In this study, we demonstrate that metformin notably attenuates Cd-evoked SGN degeneration by restoring impaired autophagy flux, as evidenced by the suppression of Cd-induced elevation of autophagy markers microtubule-associated protein 1A/1B-light chain 3-II (LC3-II) and autophagy substrate protein p62 in degenerated SGN. Blockage of autophagy flux by chloroquine abolished metformin-induced neuroprotection against Cd-induced neurotoxicity in SGN. The results of this study reveal that autophagy dysfunction is an important component of Cd-induced SGN degeneration, and metformin may be a potential protective agent for attenuating SGN degeneration following Cd exposure.

Design and Experimental Investigations of Shape and Attitude Carding System for the Wires of Micro Coreless Motor Winding.

The shape and attitude (S&A) of the electrode wire are important characteristics of micro coreless motor winding. The purpose of this paper is to present the design of a robotic micro-manipulation system for micro wire carding with arbitrary S&A, which can be used as the pretreatment system for wire micro-gripper systems. The system is based on the principle of flexible carding, and uses nylon, bristle, nanometer-silk and wool as materials for the brushing micro-manipulator. The trajectory of the brushing micro-manipulator is designed, and the S&A of the electrode wires are straightened through the combined motion mode of horizontal and vertical brushing micro-manipulators. The experimental results show that the material of the brushing micro-manipulator has a great impact on the carding quality. Nanometer-silk material is more suitable for horizontal brushing micro-manipulators, and wool material is more suitable for vertical brushing micro-manipulators. The geometric dimension of the brushing micro-manipulator also affects the carding quality. When the diameter is in the range of 1 mm, the carding effect of the horizontal brushing micro-manipulator with a length of 4.9-8 mm is better. The system can realize the automatic carding of flexible electrode wires with arbitrary S&A, and it will not damage the structure of wires in the process.

The expression profile and clinical application value of hsa_circ_0016148 in head and neck squamous cell carcinoma.

BACKGROUND: Dysregulated circular RNAs (circRNAs) are involved in human cancers and may be used as biomarkers with the potential of clinical application. However, little is known regarding the mechanism of circRNAs and their clinical application value in head and neck squamous cell carcinoma (HNSCC). METHODS: In the current study, we established the profile of circRNAs in HNSCC using microarray and then measured the expression of hsa_circ_0016148 in 137 paired HNSCC tissues by qRT-PCR technique, analyzed the relationship between hsa_circ_0016148 and clinicopathological data, and investigated its diagnostic and prognostic value. The hsa_circ_0016148-miRNA-mRNA interaction network was predicted and constructed by Cytoscape. RESULTS: Our study showed a circRNA expression profile and confirmed downregulated hsa_circ_0016148 in HNSCC tissues (p = 2.64E-35). The hsa_circ_0016148 expression is remarkably correlated with lymph node metastasis (p = 0.001) and clinical stage (p = 0.029). Then, the area under the receiver characteristic curve (AUC) was 0.912 with 92% of sensitivity and 86.9% specificity, respectively. Besides, our study demonstrated that lower-expressed hsa_circ_0016148 could independently predict poorer overall survival of HNSCC patients (hazard ratio [HR] = 0.456; 95% confidence interval [CI] = 0.265-0.784; p = 0.005). The hsa_circ_0016148-miRNA-mRNA interaction network was constructed, which included a total of nine targeted miRNAs. CONCLUSION: Taken together, our results revealed that hsa_circ_0016148 might play a critical role in HNSCC tumorigenesis and may serve as an indicator with the potential of diagnosis and prognosis for HNSCC.

The Role and Application of Salivary Exosomes in Malignant Neoplasms.

The study of salivary exosomes in malignant neoplasms has attracted widespread attention in the clinical setting. Although a variety of diagnostic and treatment approaches have been proposed, there are some limitations to their application. In recent years, the role of salivary exosomes in cancer has been increasingly studied. Salivary exosomes not only renew and regulate the biological behavior of tumor cells, such as malignant proliferation, migration, and invasion, but they also serve as ideal markers for early diagnosis of diseases and may represent an effective therapeutic target. This article reviews the current research on salivary exosomes in malignant neoplasms.

BIX-01294, a G9a inhibitor, suppresses cell proliferation by inhibiting autophagic flux in nasopharyngeal carcinoma cells.

G9a, a histone methyltransferase, has been found to be upregulated in a range of tumor tissues, and contributes to tumor growth and metastasis. However, the impact of G9a inhibition as a potential therapeutic target in nasopharyngeal carcinoma (NPC) is unclear. In the present study we aimed to investigate the anti-proliferative effect of G9a inhibition in the NPC cell lines CNE1 and CNE2, and to further elucidate the molecular mechanisms underlying these effects. The expression of G9a in NPC tumor tissues was significantly higher than that in normal nasopharyngeal tissues. The pharmacological inhibition of G9a by BIX-01294 (BIX) inhibited proliferation and induced caspase-independent apoptosis in NPC cells in vitro. Treatment with BIX induced autophagosome accumulation, which exacerbated the cytotoxic activity of BIX in NPC cells. Mechanistic studies have found that BIX impairs autophagosomes by initiating autophagy in a Beclin-1-independent way, and impairs autophagic degradation by inhibiting lysosomal cathepsin D activation, leading to lysosomal dysfunction. BIX was able to suppress tumor growth, possibly by inhibiting autophagic flux; it might therefore constitute a promising candidate for NPC therapy.

BIX-01294-enhanced chemosensitivity in nasopharyngeal carcinoma depends on autophagy-induced pyroptosis.

Nasopharyngeal carcinoma (NPC) is a common cancer in southern China and Southeast Asia. Nowadays, radiotherapy is the therapy of choice for NPC patients, and chemotherapy has been found as an alternative treatment for advanced NPC patients. However, finding novel drugs and pharmacologically therapeutic targets for NPC patients is still urgent and beneficial. Our study showed that BIX-01294 (BIX) can induce autophagic vacuoles formation and conversion of LC3B-I to LC3B-II in NPC cells in both dose- and time-dependent manners. Notably, the combination of BIX and chemotherapeutic drugs significantly decreased the cell viability and increased the lactate dehydrogenase release. Meanwhile, BIX plus cis-platinum (Cis) treatment induced pyroptosis in NPC cells as featured by cell swelling and bubble blowing from the plasma membrane, the increased frequency of annexin V and propidium iodide (PI) double-positive cells, as well as the cleavage of gasdermin E (GSDME) and caspase-3. Moreover, the deficiency of GSDME completely shifted pyroptosis to apoptosis. Furthermore, the inhibition of autophagy by chloroquine and the knockout of ATG5 gene significantly blocked the BIX-induced autophagy as well as pyroptosis in both in vitro and in vivo studies. Our data demonstrated that BIX-combined chemotherapeutic drugs could induce the Bax/caspase-3/GSDME-mediated pyroptosis through the activation of autophagy to enhance the chemosensitivity in NPC.

miR-497 may enhance the sensitivity of non-small cell lung cancer cells to gefitinib through targeting the insulin-like growth factor-1 receptor.

BACKGROUND: Recently, studies have demonstrated that microRNA-497 (miR-497) plays an important role in modulating tumor cell sensitivity to chemotherapeutic drugs; however, its role in cellular resistance to the effects of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) in treatment of non-small cell lung cancer (NSCLC) is not fully understood. In this study, we explored the potential of miR-497 in targeting the insulin-like growth factor-1 receptor (IGF-1R) signaling pathways to overcome gefitinib resistance. METHODS: A gefitinib resistant human lung adenocarcinoma A549 cell line (A549/GR) was established by the method of gefitinib mutagenesis culture. Next, the A549/GR cells were transfected with miR-497 mimics to establish an miR-497 overexpression model, designated A549/GR-miR497-mimic. MTT assay was used to assess cell resistance to gefitinib, and western blot assay was employed to evaluate alterations of IGF-1R and the AKT1 signaling pathway. RESULTS: We found that A549/GR-miR497-mimic cells (IC50 =33.76±0.97 µmol/L) were more sensitive to gefitinib than the control group (P<0.01). In addition, the expression levels of IGF-1R and phosphorylated AKT1 (p-AKT1) in A549/GR-miR497-mimic cells were reduced. CONCLUSIONS: We demonstrated that miR-497 may have the effect of reversing gefitinib resistance and increasing the sensitivity of NSCLC cells to EGFR-TKIs by inhibiting the expression of IGF-1R and reducing activation of the downstream AKT signaling pathway. Thus, miR-497 plays a vital role in the acquired resistance to EGFR-TKIs, and it may represent a potential therapeutic strategy to treat NSCLC exhibiting resistance to EGFR-TKIs.

Parapharyngeal liposarcoma: a case report.

BACKGROUND: Parapharyngeal liposarcoma is a very rare malignant tumor that often causes nonspecific clinical symptoms, such as progressive dysphagia, globus sensation and/or respiratory disturbances. The combination of radiological imaging techniques and histopathological analysis provides information for diagnosis; however, the pathogenesis is still uncertain. CASE PRESENTATION: A 30-year-old male patient presented with a pharyngeal cavity mass, which had been present for 2 years. The clinical syndrome included obstructive sleep apnea symptoms (i.e., respiratory disturbances, excessive daytime somnolence, and headache) and difficulty swallowing. The radiological examination (CT) demonstrated that there was a low-density irregular solid lesion on the posterior wall of the oropharynx and laryngopharynx, which descended to the superior mediastinum and extended to the left parapharyngeal space and sternocleidomastoid muscle. The boundaries of the lesion were clear, and the lesion's density was nonuniform. Several septations inside the lesion were observed. The CT values of the lesion at the epiglottis and the vocal folds were 11 HU and minus 30 HU, respectively. After enhanced scanning, there was no apparent enhancement of the lesion: the surrounding tissue and blood vessels were squeezed and shifted, but the neighboring sclerotin of the cervical vertebrae was not invaded. The mass was removed via a transcervical approach, resulting in a complete amelioration of the patient's symptoms. Interestingly, immunohistochemistry showed that the tumor cells expressed members of the B7 superfamily, including B7-H1, B7-DC and B7-H3. In addition, the expression of TIM-containing molecules, including TIM-3 and TIM-4, was observed. CONCLUSIONS: CT and MRI demonstrated that the mass was a parapharyngeal liposarcoma. Furthermore, carcinoma-associated B7 and TIM-containing molecules were observed in the tissue, indicating that these molecules are most likely active in the pathogenesis of this disease. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1907794973876202.