Insights into the heterogeneity of the tumor microenvironment in lung adenocarcinoma and squamous carcinoma through single-cell transcriptomic analysis: Implications for distinct immunotherapy outcomes.

BACKGROUND: Immune checkpoint blockade has emerged as a key strategy to the therapy landscape of non-small cell lung cancer (NSCLC). However, notable differences in immunotherapeutic outcomes exist between the two primary NSCLC subtypes: lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). This disparity may stem from the tumor immune microenvironment's heterogeneity at the transcriptome level. METHODS: By integrative analysis of transcriptomic characterization of 38 NSCLC patients by single-cell RNA sequencing, the present study revealed a distinct tumor microenvironment (TME) between LUAD and LUSC, with relevant results further confirmed in bulk transcriptomic and multiplex immunofluorescence (mIF) validation cohort of neoadjuvant immunotherapy patients. RESULTS: LUAD exhibited a more active immune microenvironment compared to LUSC. This included highly expression of HLA I/II in cancer cells, reinforced antigen presentation potential of dendritic cells and enhanced cytotoxic activity observed in T/NK cells. In LUSC, cancer cells highly expressed genes belonging to the aldo-keto reductases, glutathione S-transferases and aldehyde dehydrogenase family, negatively correlating with immunotherapy outcomes in the validation cohort of our center. Further analysis revealed elevated infiltrated cancer-associated fibroblasts (CAFs) in LUSC, which was corroborated in The Cancer Genome Atlas cohort. Corresponding increased infiltration of ADH1B+ CAFs in major pathologic response (MPR) patients and the higher presence of FAP+ CAFs in non-MPR patients were demonstrated by multiplex mIF. Moreover, upregulating immunosuppressive extracellular matrix remodeling was identified in LUSC. CONCLUSIONS: These comprehensive analyses advance the understanding of the differences in TME between LUAD and LUSC, offering insights for patient selection and developing subtype-specific treatment strategies.

Down-regulated HHLA2 enhances neoadjuvant immunotherapy efficacy in patients with non-small cell lung cancer (NSCLC) with chronic obstructive pulmonary disease (COPD).

BACKGROUND: Emerging data suggested a favorable outcome in advanced non-small cell lung cancer (NSCLC) with chronic obstructive pulmonary disease (COPD) patients treated by immunotherapy. The objective of this study was to investigate the effectiveness of neoadjuvant immunotherapy among NSCLC with COPD versus NSCLC without COPD and explore the potential mechanistic links. PATIENTS AND METHODS: Patients with NSCLC receiving neoadjuvant immunotherapy and surgery at Shanghai Pulmonary Hospital between November 2020 and January 2023 were reviewed. The assessment of neoadjuvant immunotherapy's effectiveness was conducted based on the major pathologic response (MPR). The gene expression profile was investigated by RNA sequencing data. Immune cell proportions were examined using flow cytometry. The association between gene expression, immune cells, and pathologic response was validated by immunohistochemistry and single-cell data. RESULTS: A total of 230 NSCLC patients who received neoadjuvant immunotherapy were analyzed, including 60 (26.1%) with COPD. Multivariate logistic regression demonstrated that COPD was a predictor for MPR after neoadjuvant immunotherapy [odds ratio (OR), 2.490; 95% confidence interval (CI), 1.295-4.912; P = 0.007]. NSCLC with COPD showed a down-regulation of HERV-H LTR-associating protein 2 (HHLA2), which was an immune checkpoint molecule, and the HHLA2low group demonstrated the enrichment of CD8+CD103+ tissue-resident memory T cells (TRM) compared to the HHLA2high group (11.9% vs. 4.2%, P = 0.013). Single-cell analysis revealed TRM enrichment in the MPR group. Similarly, NSCLC with COPD exhibited a higher proportion of CD8+CD103+TRM compared to NSCLC without COPD (11.9% vs. 4.6%, P = 0.040). CONCLUSIONS: The study identified NSCLC with COPD as a favorable lung cancer type for neoadjuvant immunotherapy, offering a new perspective on the multimodality treatment of this patient population. Down-regulated HHLA2 in NSCLC with COPD might improve the MPR rate to neoadjuvant immunotherapy owing to the enrichment of CD8+CD103+TRM. TRIAL REGISTRATION: Approval for the collection and utilization of clinical samples was granted by the Ethics Committee of Shanghai Pulmonary Hospital (Approval number: K23-228).

Stratifin promotes renal dysfunction in ischemic and nephrotoxic AKI mouse models via enhancing RIPK3-mediated necroptosis.

Stratifin (SFN) is a member of the 14-3-3 family of highly conserved soluble acidic proteins, which regulates a variety of cellular activities such as cell cycle, cell growth and development, cell survival and death, and gene transcription. Acute kidney injury (AKI) is prevalent disorder characterized by inflammatory response, oxidative stress, and programmed cell death in renal tubular epithelial cells, but there is still a lack of effective therapeutic target for AKI. In this study, we investigated the role of SFN in AKI and the underlying mechanisms. We established ischemic and nephrotoxic AKI mouse models caused by ischemia-reperfusion (I/R) and cisplatin, respectively. We conducted proteomic and immunohistochemical analyses and found that SFN expression levels were significantly increased in AKI patients, cisplatin- or I/R-induced AKI mice. In cisplatin- or hypoxia/reoxygenation (H/R)-treated human proximal tubule epithelial cells (HK2), we showed that knockdown of SFN significantly reduced the expression of kidney injury marker Kim-1, attenuated programmed cell death and inflammatory response. Knockdown of SFN also significantly alleviated the decline of renal function and histological damage in cisplatin-caused AKI mice in vivo. We further revealed that SFN bound to RIPK3, a key signaling modulator in necroptosis, to induce necroptosis and the subsequent inflammation in cisplatin- or H/R-treated HK2 cells. Overexpression of SFN increased Kim-1 protein levels in cisplatin-treated MTEC cells, which was suppressed by RIPK3 knockout. Taken together, our results demonstrate that SFN that enhances cisplatin- or I/R-caused programmed cell death and inflammation via interacting with RIPK3 may serve as a promising therapeutic target for AKI treatment.

Anatomical localization of horizontal segment of the petrous internal carotid artery in transnasal endoscopic skull base surgery.

OBJECTIVE: To discuss the localization of horizontal segment of petrous internal carotid artery in transnasal endoscopic skull base surgery, and to provide anatomical data for clinical surgery. METHODS: The horizontal segment of the petrous internal carotid artery of 5 adult cadaveric heads were exposed by endoscopic transnasal and microscopic open approaches respectively, and the relevant data and images were measured and collected. RESULTS: The medial wall of the foramen spinosum is the lateral wall of the isthmus of the eustachian tube, and the thickness of the bone is 0.5 ± 0.2 mm. The medial wall of the isthmus of the eustachian tube is the lateral wall of the posterior genu of the carotid canal and the thickness of the bone is 0.2 ± 0.1 mm. The vidian nerve originates from the anterior genu of the petrous internal carotid artery. The distance from the base of vidian nerve to the isthmus of eustachian tube is 19.2 ± 2.8 mm. CONCLUSION: The foramen spinosum is the landmark of isthmus of the Eustachian tube. The isthmus of the Eustachian tube is the landmark of the posterior genu of the internal carotid artery. The line between the base of the vidian nerve and the isthmus of the Eustachian tube ioks the landmark of horizontal segment of the petrous internal carotid artery.

The anatomy of the parapharyngeal segment of the internal carotid artery for endoscopic endonasal approach.

Injury to the internal carotid artery (ICA) is a life-threatening complication of endoscopic endonasal approaches. The objective of this study is to illustrate the detail anatomy of the parapharyngeal segment of the ICA (PPICA) to safe endoscopic endonasal surgery. The anatomical dissection was performed in 10 cadaveric specimens and several crucial anatomical landmarks were identified and measured. In addition, 50 dry skulls were studied to further assess the relationship between the pharyngeal tubercle and carotid foramen. From the endoscopic endonasal perspective, in the median plane, the pharyngeal tubercle and the carotid foramen on both sides were located on a line. The average distance between the pharyngeal tubercle and anterior border of the external orifice of the carotid canal was measured as 25.2 ± 3.2 mm. In the paramedian plane, the PPICA was located between the levator veli palatini muscle (LVPM) and the stylopharyngeal muscle (SPM) in upper parapharyngeal space in all specimens, and the distance from the posterior border of the LVPM to the anterior border of the SPM was recorded as 15.1 ± 2.8 mm at the level of the carotid foramen. The distance from the attachment of the LVPM to the anterior border of the external orifice of the carotid canal was about 5.1 ± 0.2 mm. The fully developed stylopharyngeal fascia (SPhF) was observed in 10 cases, and the PPICA was always anteriorly enclosed by and adhered to the SPhF.

MAIT cells are associated with responsiveness to neoadjuvant immunotherapy in COPD-associated NSCLC.

BACKGROUND: Patients with non-small cell lung cancer (NSCLC) and chronic obstructive pulmonary disease (COPD) experience worse clinical outcomes but respond better to immunotherapy than patients with NSCLC without COPD. Mucosal-associated invariant T (MAIT) cells, a versatile population of innate immune T lymphocytes, have a crucial function in the response to infection and tumors. This study investigated the distribution of MAIT cells in COPD-associated NSCLC and their involvement in the immune response. METHODS: Flow cytometry, immunohistochemistry, and immunofluorescence were performed on tissue samples of patients with NSCLC, with or without COPD, treated with or without anti-programmed death 1 (PD1) immunotherapy. MAIT cells were stimulated with 5-OP-RU using a mouse subcutaneous tumor model. RESULTS: Tumors contained significantly more MAIT cells than paraneoplastic tissues, and CD8+ MAIT cells accounted for more than 90% of these cells. Patients with NSCLC and COPD had higher CD8+ MAIT cell counts than those with NSCLC without COPD. Additionally, patients with NSCLC and COPD displayed reduced expression of the activation marker, CD69, and functional markers, granzyme B (GZMB) and interferon γ (IFNγ), and higher expression of the immune exhaustion marker, PD1. Among patients who received immunotherapy, the proportion with a complete or partial response was higher in those with COPD than in those without COPD. In patients with NSCLC and COPD, the major pathologic response (MPR) group had higher MAIT levels than those in the no major pathologic response (NPR) group. In the mouse subcutaneous tumor model stimulation of MAIT cells using 5-OP-RU enhanced the antitumor effects of anti-PD1. CONCLUSIONS: In patients with NSCLC and COPD, response to immunotherapy is associated with accumulation of CD8+ MAIT cells showing immune exhaustion. These findings may contribute to innovative approaches for immunotherapy targeting CD8+ MAIT cells.

Modified Skin Incision and Location of Burr-Hole Surgery via a Retrosigmoid Approach: An Anatomical Study.

Objective This study aims to reduce the tissue damage during craniotomy with retrosigmoid approach. A modified sickle-shaped skin incision was developed, and a new burr-hole positioning method was proposed. Methods Five adult cadaveric heads (10 sides) were used in this study. The sickle-shaped skin incision was performed during craniotomy. The nerves, blood vessels, and muscles were observed and measured under a microscope. Additionally, 62 dry adult skull specimens (left sided, n = 35; right sided, n = 27) were used to measure the distance between the most commonly used locating point (asterion [Ast] point) and the posteroinferior point of the transverse sigmoid sinus junction (PSTS) (Ast-PSTS), as well as the distance between the new locating O point and the PSTS (O-PSTS). Then, the reliability of the new locating O point was validated on the same five adult cadaveric heads (10 sides) used for the sickle-shaped skin incision. Results The sickle-shaped skin incision reduced the damage to the occipital nerves, blood vessels, and muscles during the surgery via a retrosigmoid approach. The dispersion and variability of O-PSTS were smaller than those of Ast-PSTS. Conclusion The sickle-shaped skin incision of the retrosigmoid approach can reduce the tissue damage and can completely expose the structures in the cerebellopontine angle. The modified O point is a more reliable locating point for a burr-hole surgery than the Ast point.

The endoscopic prelacrimal recess approach to the paramedian middle cranial base: An anatomical study.

BACKGROUND: Endoscopic endonasal approach to paramedian cranial base implies sacrifice of the nasal structures. OBJECTIVE: The present study aimed to illustrate the anatomy and provide critical anatomical landmarks for the endoscopic prelacrimal recess approach (PLRA) to the paramedian middle cranial base. METHODS: Anatomical dissections were performed in 10 cadaveric specimens. RESULTS: Successful access to the paramedian middle cranial base was achieved in all dissections via the PLRA with the removal of the pterygoid process. For the dissection of the infratemporal fossa and pterygopalatine fossa, the buccal nerve and infraorbital neurovascular bundle can serve as important anatomic landmarks to identify the detailed structures. In the upper parapharyngeal space, the stylopharyngeal aponeurosis can present as anatomical barriers to protect the parapharyngeal segment of the internal carotid artery (PPICA); while the levator veli palatini muscle can be considered as a landmark to locate the PPICA. For the dissection of the Eustachian tube (ET), the isthmus of the ET and ET sulcus can serve as useful landmarks to identify the posterior genu of the ICA and horizontal segment of the petrous ICA respectively. CONCLUSION: The PLRA to the paramedian middle cranial base is anatomically feasible and can facilitate preservation of the integrity of nasal structures. The buccal nerve, infraorbital neurovascular bundle, levator veli palatini muscle, stylopharyngeal aponeurosis, the isthmus of the ET, and ET sulcus can serve as critical anatomic landmarks in their respective region and may facilitate the application of this approach.

Transmastoid Trautman's Triangle Combined Low Retrosigmoid Approach for Foramen Magnum Meningiomas: Surgical Anatomy and Technical Note.

Objective This study was aimed to assess the potential of utilizing a transmastoid Trautman's triangle combined low retrosigmoid approach for ventral and ventrolateral foramen magnum meningiomas (FMMs) surgical treatment. Methods We simulated this transmastoid Trautman's triangle combined low retrosigmoid approach using five adult cadaveric heads to explore the associated anatomy in a step-by-step fashion, taking pictures of key positions as appropriate. We then employed this approach in a single overweight patient with a short neck who was suffering from large ventral FMMs and cerebellar tonsillar herniation. Results Through cadaver studies, we were able to confirm that this transmastoid Trautman's triangle combined with low retrosigmoid approach achieves satisfactory cranial nerve and vasculature visualization while also offering a wide view of the whole of the ventrolateral medulla oblongata. We, additionally, have successfully employed this approach to treat a single patient suffering from large ventral FMMs with cerebellar tonsillar herniation. Conclusion This transmastoid Trautman's triangle combined low retrosigmoid approach may represent a complement to treatment strategies for ventral and ventrolateral FMMs, particularly in patients with the potential for limited surgical positioning due to their being overweight, having a short neck and suffering from cerebellar tonsillar herniation.

Knockdown of the long noncoding RNA XIST suppresses glioma progression by upregulating miR-204-5p.

Background: Gliomas are the most prevalent primary malignant tumors of the central nervous system. Our previous study showed that miR-204-5p is a tumor suppressor gene in glioma. Bioinformatic analyses suggest that long noncoding RNA (lncRNA) X-inactive specific transcript (XIST) is a potential target gene of miR-204-5p. Methods: We analyzed the expression of XIST and miR-204-5p in glioma tissues and the correlation with glioma grade. A series of in vitro experiments were carried out to elucidate the role of XIST in glioma progression. A mouse xenograft model was established to detect whether knockdown of XIST can inhibit glioma growth. A luciferase assay was performed to determine whether XIST can bind to miR-204-5p and the binding specificity. Cells stably expressing shXIST or shNC were transfected with anti-miR-204-5p or anti-miR-204-5p-NC to evaluate whether XIST mediates the tumor-suppressive effects of miR-204-5p. Results: XIST was upregulated in glioma tissues compared with normal brain tissues (NBTs), while miR-204-5p expression was significantly decreased in glioma tissues compared with NBTs. Both XIST and miR-204-5p expression levels were clearly related to glioma grade, and the expression of XIST was obviously negatively correlated with miR-204-5p expression. Knockdown of XIST inhibited glioma cell proliferation, migration, and invasion, promoted apoptosis of glioma cells, inhibited tumor growth and increased the survival time in nude mice. miR-204-5p could directly bind to XIST and negatively regulate XIST expression. XIST mediated glioma progression by targeting miR-204-5p in glioma cells. XIST crosstalk with miR-204-5p regulated Bcl-2 expression to promote apoptosis. Conclusion: Our results provide evidence that XIST, miR-204-5p and Bcl-2 form a regulatory axis that controls glioma progression and can serve as a potential therapeutic target for glioma.

A Microanatomical Study of the Far Lateral Approach.

OBJECTIVE: The far-lateral approach (FLA) remains a challenge for neurosurgeons due to the complex anatomy of this region, especially in patients with anatomical variations. There is therefore an urgent need for better quantitative knowledge of the microsurgical anatomy of the FLA. METHODS: The study was performed using the dried skulls and atlas vertebrae of 50 Chinese adults, in which significant clinical parameters were measured. We further used 12 cadaveric heads to simulate the FLA to explore the step-by-step anatomy entailed by this procedure, enabling us to obtain key images and related information. RESULTS: Limited to hypoglossal canal, the occipital condyle posterior was abraded by roughly 10 mm, which provided good exposure to the ventral front of the foramen magnum. When occipital artery exits the occipital groove, the mean diameter was 2.20 mm. The average occipital artery suboccipital segment length was 65.26 mm. The posterior spinal artery (PSA) and posterior inferior cerebellar artery (PICA) generally originated from the fourth vertebral artery segment intradurally, and the mean distances from the PSA and PICA to the dural entry point of the vertebral artery were 2.62 mm and 8.71 mm, respectively. The incidence of PSA and PICA arising from the third vertebral artery segment was 16.67% and 4.17%, respectively. CONCLUSIONS: Understanding the important anatomic structures of the CVJ region and developing improved knowledge of the microsurgical anatomy of the FLA offer an opportunity to ensure safe exposure and treatment of lesions in the ventral and ventrolateral regions of the CVJ.