Evaluation of the antitumor effect of neoantigen peptide vaccines derived from the translatome of lung cancer.

Emerging evidence suggests that tumor-specific neoantigens are ideal targets for cancer immunotherapy. However, how to predict tumor neoantigens based on translatome data remains obscure. Through the extraction of ribosome-nascent chain complexes (RNCs) from LLC cells, followed by RNC-mRNA extraction, RNC-mRNA sequencing, and comprehensive bioinformatic analysis, we successfully identified proteins undergoing translatome and exhibiting mutations in the cells. Subsequently, novel antigens identification was analyzed by the interaction between their high affinity and the Major Histocompatibility Complex (MHC). Neoantigens immunogenicity was analyzed by enzyme-linked immunospot assay (ELISpot). Finally, in vivo experiments in mice were conducted to evaluate the antitumor effects of translatome-derived neoantigen peptides on lung cancer. The results showed that ten neoantigen peptides were identified and synthesized by translatome data from LLC cells; 8 out of the 10 neoantigens had strong immunogenicity. The neoantigen peptide vaccine group exhibited significant tumor growth inhibition effect. In conclusion, neoantigen peptide vaccine derived from the translatome of lung cancer exhibited significant tumor growth inhibition effect.

circUBAP2 exacerbates malignant capabilities of NSCLC by targeting KLF4 through miR-3182 modulation.

Chemo-resistance and refractoriness remain challenges for Non-small cell lung cancer (NSCLC) patients and the underlying molecular mechanisms haven't been fully explained. In this study, we investigated the influence of circUBAP2 on the NSCLC tumor cells. This study might provide novel therapeutic targets for NSCLC treatment. Clinical samples and NSCLC cell lines were used to investigate circUBAP2 expressions and their impact on tumor cell chemo-resistance. CCK8 and transwell assays were conducted to explore the differences of NSCLC tumor proliferation and migration capabilities affected by circUBAP2. Dual-luciferase reporter gene assay was performed to explore the detailed molecular mechanism of circUBAP2 regulation network. circUBAP2 exhibited significantly elevated average level in our clinical samples of NSCLC, compared with normal tissues. CircUBAP2 level was positively correlated with disease stage and metastatic status. circUBAP2 significantly enhanced the migration, proliferation and chemo-resistance of NSCLC cell lines. Further experiments indicated that circUBAP2 promoted malignant biological behavior of NSCLC tumor cells by targeting KLF4 through modulating miR-3182 expression. Our study demonstrated for the first time that circUBAP2 played an important role exacerbating malignant capabilities of NSCLC. circUBAP2-miR3182-KLF4 regulative network demonstrated in this study could be a novel therapeutic target for future NSCLC treatment.

CircMED13L_012 promotes lung adenocarcinoma progression by upregulation of MAPK8 mediated by miR-433-3p.

BACKGROUND: Metastasis and disease refractoriness remain as major challenges for non-small cell lung cancer (NSCLC) treatment and understanding the underlying molecular mechanisms is of scientific and clinical value. Therefore, in this study, we aimed to explore the effects of circMED13L_012 on the proliferation, migration, invasion and drug-resistance of NSCLC tumor cells. METHODS: In this study, we utilized clinical samples and NSCLC cell lines to explore the association between circMED13L_012 expressions and tumor cell metastasis and chemo resistance. CCK8 and transwell assay were conducted to explore the impact of circMED13_012 on NSCLC tumor proliferation and migrative capabilities. Dual-luciferase reporter gene assay was conducted to validate the circMED13L_012 interaction network. RESULTS: Our results demonstrated that circMED13L_012 exhibited significantly elevated average level in our clinical samples of NSCLC, compared with normal tissues. circMED13L_012 level was positively correlated with disease stage and metastatic status. Increased circMED13L_012 expression was associated with the enhanced migration, proliferation and chemo resistance of NSCLC cell lines. Further experiments indicated that circMED13L_012 promoted malignant behavior of NSCLC tumor cells by targeting MAPK8 through modulation miR-433-3p expression. CONCLUSIONS: Our study for the first time demonstrated that circMED13L_012-miR-433-3p-MAPK8 axis played important role for NSCLC pathogenesis, which could be potential therapeutic target for the development of future NSCLC treatment.

E2F1-induced ferritin heavy chain 1 pseudogene 3 (FTH1P3) accelerates non-small cell lung cancer gefitinib resistance.

Long noncoding RNAs (lncRNAs) have been identified to be critical regulator for various human diseases and emerging evidence illustrate the essential function of lncRNAs in the non-small cell lung cancer (NSCLC). Here, our research team tried to identify the roles of lncRNA ferritin heavy chain 1 pseudogene 3 (FTH1P3) in the NSCLC, as well as its molecular mechanism. LncRNA microarray analysis revealed that ferritin heavy chain 1 pseudogene 3 (FTH1P3) was up-regulated in the gefitinib-resistant cells (PC9/GR). Clinically, lncRNA FTH1P3 high-expression was closely correlated with NSCLC patients' unfavorable prognosis. Gain and loss of functional experiments revealed that FTH1P3 promoted the proliferation and invasion of NSCLC cells in vitro, and FTH1P3 knockdown repressed the tumor growth in vivo. Mechanistically, transcription factor E2F1 accelerated the transcription of FTH1P3. RNA immunoprecipitation and chromatin immunoprecipitation experiments showed that FTH1P3 can recruit lysine-specific demethylase 1 (LSD1) and epigenetically repress the TIMP3, thereby accelerating the tumorigenesis of NSCLC. In summary, these findings suggest that FTH1P3 plays a critical role in the gefitinib resistance and progression of NSCLC, providing a potential novel prognostic marker for NSCLC.

Effects of the tumor suppressor PTEN on the pathogenesis of idiopathic pulmonary fibrosis in Chinese patients.

Idiopathic pulmonary fibrosis (IPF) is characterized by progressive interstitial fibrosis, and is associated with a fatal outcome. The critical pathological mechanisms underlying IPF are largely unknown; however, accumulating evidence has indicated similarities between IPF and cancer. Therefore, the present study examined the expression levels of the tumor suppressor phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in Chinese patients with IPF, using an enzyme­linked immunosorbent assay. To determine the effects of PTEN on the development of pulmonary fibrosis, PTEN was overexpressed in transforming growth factor (TGF)­ß1­treated human embryonic lung fibroblasts (HFL­I cells). The serum levels of PTEN were significantly lower in 42 patients with IPF, as compared with in the healthy controls. In addition, PTEN overexpression enhanced apoptosis, and suppressed basal levels of proliferation and migration in fibroblasts. Notably, PTEN was able to specifically inhibit TGF­ß1­induced proliferation and migration of the cells. Overexpression of PTEN also suppressed phosphorylation of Akt and Smad3, and decreased the expression levels of numerous proteins with critical roles in TGF­ß1­induced fibrosis, including α­smooth muscle actin, matrix metalloproteinase (MMP)­2 and MMP­9. These results indicated that PTEN may inhibit TGF­ß1­mediated myofibroblast differentiation of fibroblasts by attenuating signaling via the phosphatidylinositol 3­kinase/Akt and TGF­ß/Smad3 pathways.

CYP2J2 and EETs Protect against Oxidative Stress and Apoptosis in Vivo and in Vitro Following Lung Ischemia/Reperfusion.

BACKGROUND: Cytochrome P450 epoxygenase 2J2 (CYP2J2) metabolizes arachidonic acids to epoxyeicosatrienoic acids (EETs). EETs exert various biological effects, including anti-inflammatory, anti-apoptotic, pro-proliferation, pro-angiogenesis, anti-oxidation, and anti-fibrosis effects. However, little is known about the role of CYP2J2 and EETs in lung ischemia/reperfusion injury. In this study, we examined the effects of exogenous EETs or CYP2J2 overexpression on lung ischemia/reperfusion injury in vivo and in vitro. METHODS AND RESULTS: CYP2J2 gene was stably transfected into rat lungs via pcDNA3.1-CYP2J2 plasmid delivery, resulting in increased EETs levels in the serum and lung. A rat model of lung ischemia/reperfusion injury was developed by clamping the left lung hilum for 1 hour, followed by reperfusion for 2 hours. We found that CYP2J2 overexpression markedly decreased the levels of oxidative stress and cell apoptosis in lung tissues induced by ischemia/reperfusion. Moreover, we observed that exogenous EETs, or CYP2J2 overexpression, enhanced cell viability, decreased intracellular reactive oxygen species (ROS) generation, inhibited mitochondrial dysfunction, and attenuated several apoptotic signaling events in a human pulmonary artery endothelial cells (HPAECs)-based anoxia/reoxygenation model. These apoptotic events included activation of NADPH oxidase, collapse of mitochondrial transmembrane potential, and activation of pro-apoptotic proteins and caspase-3. These effects were mediated, at least partially, by the PI3K/Akt signaling pathway. CONCLUSION: These results reveal that CYP2J2 overexpression and exogenous EETs can protect against oxidative stress and apoptosis following lung ischemia/reperfusion in vivo and in vitro, suggesting that increasing the level of EETs may be a novel promising strategy to prevent and treat lung ischemia/reperfusion injury.

[Peripheral primitive neuroectodermal tumor of trachea: a case report and review of literatures].

OBJECTIVE: To improve the understanding of tracheal peripheral primitive neuroectodermal tumor (PNET). METHODS: A case of tracheal PNET diagnosed in July 2010 was reported and the related literatures were reviewed. The literature review was carried out respectively with "primitive neuroectodermal tumor", "peripheral" as the search terms in Wanfang med online and PubMed database by September 2011. RESULTS: A case of 63 year-old female patient, who had been misdiagnosed as having chronic pharyngitis, chronic bronchitis and bronchial asthma, was admitted to the hospital because of cough and sputum production for 50 days, and anhelation for 1 month. After admission, the chest computerized tomography showed a space-occupying lesion in the middle of the trachea. Bronchoscopy showed a pedicle neoplasm 4 cm under the subglottic, with integral capsule, smooth surface and rich vascellum. Subsequently, tumor resection under bronchoscope was performed. Pathology report after operation showed infiltration of flake small round malignant cells under bronchial mucosa. Immunohistochemistry showed CD(99)(+), Syn(+) and S-100(+). EWS-FLI-1 fusion transcript was detected by RT-PCR. Accordingly, it was diagnosed as PNET. The symptoms of cough and anhelation were disappeared after operation. So far, there was no local recurrence and distant metastasis with 14 months follow-up. A total of 111 literatures were received in Pubmed, including one of prospective study, one of review, 22 of retrospective study and 87 of case report. Forty literatures and 187 cases in all were received in Wanfang Med Online, including 24 of retrospective study and 16 of case report. But, there were no reports about tracheal PNET. CONCLUSIONS: PNET can occur in the trachea and is easy to be misdiagnosed. To make a definite diagnosis, histopathology and immunohistochemistry are needed and detection of EWS-FLI-1 fusion transcript is a reliable marker for molecular diagnosis. The tracheal pPNET may be different with the pPNETs in other parts, and has a lower-grade invasion and less distant metastasis.

[Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease) of the airway: a case report and review of the literature].

OBJECTIVE: To improve the awareness of sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease) involving the airways. METHODS: The clinical presentations, endoscopic findings in the airways, pathological characteristics, and diagnosis and treatment of a case of Rosai-Dorfman disease was reported, and related literatures were reviewed. RESULTS: A 60-year-old female patient was admitted to this hospital because of recurrent wheezing for 18 months and aggravated for 1 month on March 6, 2007. The diagnosis of "bronchial asthma" had been made and oral prednisolone and inhaled budesonide resulted in symptom improvement. One month ago, she had wheezing again with inspiratory dyspnea, which was more obvious at recumbent position.She had been found to have high blood pressure and left adrenal adenoma 23 years ago, and as the diagnosis of "primary aldosteronism" was made but underwent no surgery. Left parotid gland tumor and left submandibular lymph nodes had been found, and surgical resection implemented in 1999. Lacrimal gland tumor resection of her eyes had been performed in 2000. Multiple subcutaneous nodules, rising and disappearing spontaneously, had been demonstrated in 2001. After admission, physical examination revealed nodules of 3.0 cm x 2.0 cm in her left submandibular area, and soybean sized nodules at both arms, back, chest, abdomen, buttocks and thighs. Chest CT scan and tracheal reconstruction showed that there were multiple nodules in the tracheal wall with narrow lumen, with no obvious enlargement of mediastinal lymph nodes. Lymph node biopsy showed faintly stained areas and the formation of plasma cells and lymphocytes of the deeply stained area, presenting as a sinus-like structure, and plasma cells and lymphocytes were engulfed in the plasma of the histiocytes, consistent with the diagnosis of Rosai-Dorfman disease. CONCLUSIONS: Rosai-Dorfman disease involving the airway was a rare disease often misdiagnosed. Bronchoscopy was very helpful for the diagnosis. Histiocytosis with phagocytosis of plasma cells and lymphocytes was the pathological feature, and immunohistochemical staining positive for S100 protein and CD(68) was suggestive of the diagnosis. Surgical resection combined with corticosteroids or radiotherapy was effective treatment of the airway diseases.