A phase II study of efficacy and safety of the MEK inhibitor tunlametinib in patients with advanced NRAS-mutant melanoma.

BACKGROUND: NRAS-mutant melanoma is an aggressive subtype with poor prognosis; however, there is no approved targeted therapy to date worldwide. METHODS: We conducted a multicenter, single-arm, phase II, pivotal registrational study that evaluated the efficacy and safety of the MEK inhibitor tunlametinib in patients with unresectable, stage III/IV, NRAS-mutant melanoma (NCT05217303). The primary endpoint was objective response rate (ORR) assessed by independent radiological review committee (IRRC) per Response Evaluation Criteria in Solid Tumors (RECIST) v1.1. The secondary endpoints included progression-free survival (PFS), disease control rate (DCR), duration of response(DOR), overall survival (OS) and safety. FINDINGS: Between November 2, 2020 and February 11, 2022, a total of 100 patients were enrolled. All (n = 100) patients received at least one dose of tunlametinib (safety analysis set [SAS]) and 95 had central laboratory-confirmed NRAS mutations (full analysis set [FAS]). In the FAS, NRAS mutations were observed at Q61 (78.9%), G12 (15.8%) and G13 (5.3%). The IRRC-assessed ORR was 35.8%, with a median DOR of 6.1 months. The median PFS was 4.2 months, DCR was 72.6% and median OS was 13.7 months. Subgroup analysis showed that in patients who had previously received immunotherapy, the ORR was 40.6%. No treatment-related deaths occurred. INTERPRETATION: Tunlametinib showed promising antitumor activity with a manageable safety profile in patients with advanced NRAS-mutant melanoma, including those who had prior exposure to immunotherapy. The findings warrant further validation in a randomized clinical trial.

Nonossifying fibroma of the lower femur with genu valgum: a case report.

BACKGROUND: Nonossifying fibroma is common in children and adolescents, and nonossifying fibroma with genu valgum is rare in the clinic. This article evaluated the effectiveness of treatment in a case of nonossifying fibroma of the lower femur with genu valgum. CASE PRESENTATION: A 16-year-old girl complained of pain in the lower part of her right thigh for one year. She was diagnosed as non ossifying fibroma of the right femur with secondary valgus deformity of the right knee, and was treated in our hospital. We performed curettage, bone grafting and internal fixation,and corrected the valgum deformity at the same time. The patient's incision healed well, the pain was disappeared, and the mechanical axis of lower limbs was corrected. No tumor recurrence was found on X- ray examination one year after operation, and the fracture end was healed. The patient could walk normally, and she was satisfied with her limb function. CONCLUSION: Nonossifying fibroma with genu valgum is rare in the clinic. The patient was satisfied with our treatment, which achieved a good curative effect.

Genome-wide identification of alternative splicing and splicing regulated in immune infiltration in osteosarcoma patients.

Background: Osteosarcoma typically occurs in adolescents, and the survival rate of patients with metastatic and recurrent osteosarcoma remains low. Abnormal regulation of alternative splicing is associated with the development of osteosarcoma. However, there is no genome-wide analysis of the function and regulatory mechanisms of aberrant alternative splicing associated with osteosarcoma. Methods: Published transcriptome data on osteosarcoma (GSE126209) derived from osteosarcoma patient tissue were downloaded. Gene expression profiling by high-throughput sequencing was performed on 9 normal samples and 10 tumor samples for genome-wide identification of osteosarcoma-related alternative splicing events. The potential function of osteosarcoma-associated alternative splicing events was examined by immune infiltration and correlation analysis. Regulation of aberrantly expressed RNA-binding proteins (RBPs) related to alternative splicing in osteosarcoma was clarified by co-expression analysis. Results: A total of 63 alternative splicing events, which are highly credible and dominant, were identified. GO enrichment analysis indicated that alternative splicing may be closely related to the immune response process. Immune infiltration analysis showed significant changes in the percentages of CD8 T cells, resting memory CD4 T cells, activated memory CD4 T cells, monocytes, resting dendritic cells, and activated mast cells in tumors compared to normal tissues, indicating the involvement of these immune cell types in the occurrence of osteosarcoma. Moreover, the analysis identified alternative splicing events that were co-altered with resting memory CD4 T cells, resting dendritic cells, and activated mast cells, events that may be associated with regulation of the osteosarcoma immune microenvironment. In addition, a co-regulatory network (RBP-RAS-immune) of osteosarcoma-associated RBPs with aberrant alternative splicing and altered immune cells was established. These RBPs include NOP58, FAM120C, DYNC1H1, TRAP1, and LMNA, which may serve as molecular targets for osteosarcoma immune regulation. Conclusion: These findings allow us to further understand the causes of osteosarcoma development and provide a new research direction for osteosarcoma immunotherapy or targeted therapy.

MicroRNA-215-5p promotes proliferation, invasion, and inhibits apoptosis in liposarcoma cells by targeting MDM2.

BACKGROUND: Liposarcoma (LPS) is one of the most common soft tissue malignancies in adults, and it is characterized by dysregulation of multiple signaling pathways, including MDM2 proto-oncogene (MDM2) amplification. MicroRNA (miRNA) regulates gene expression through incomplete complementary pairing with the 3' untranslated region of mRNAs involved in tumor progression. METHODS: In this study, bioinformatics analysis, RT-qPCR, dual-luciferase reporter gene, MTT, flow cytometry, cell scratches, chamber migration, colony formation, FISH, WB, and CCK8 were used. RESULTS: RT-qPCR showed that the expression of MDM2 was increased when miR-215-5p was overexpressed compared with the control group. The dual-luciferase reporter gene showed that the Renilla ratio firefly fluorescence intensity was decreased in the overexpression group compared with the control group. Cell phenotype experiments revealed that the overexpression group had increased cell proliferation rate, increased apoptosis rate, increased colony formation rate, increased cell healing area ratio, and increased number of cell invasions. FISH revealed increased MDM2 expression in the overexpression group. WB suggested decreased Bax expression, increased PCNA, Bcl-2, and MDM2 expression, and decreased P53 and P21 expression in the overexpression group. CONCLUSIONS: In this study, we suggest that miR-215-5p can target and promote MDM2 expression, promote the proliferation and invasion of LPS cells SW-872, and inhibit apoptosis.Targeting miR-215-5p may be a novel therapeutic strategy for the treatment of LPS.

Follow-up of biological reconstruction of epiphysis preserving osteosarcoma around the knee in children: A retrospective cohort study.

Limb salvage treatment for malignant bone tumors in children includes prosthetic and biological reconstruction. Early function following prosthesis reconstruction is satisfactory; however, there are several complications. Biological reconstruction is another way to treat bone defects. We evaluated the effectiveness of reconstruction of bone defects by liquid nitrogen inactivation of autologous bone with preserving epiphysis in 5 cases of periarticular osteosarcoma of the knee. We retrospectively selected 5 patients with articular osteosarcoma of the knee who underwent epiphyseal-preserving biological reconstruction in our department between January 2019 and January 2020. Femur involvement occurred in 2 cases and tibia involvement occurred in 3 cases, with an average defect of 18 cm (12-30 cm). The 2 patients with femur involvement were treated with inactivated autologous bone by liquid nitrogen with vascularized fibula transplantation. Among the patients with tibia involvement, 2 were treated with inactivated autologous bone with ipsilateral vascularized fibula transplantation and 1 was treated with autologous inactivated bone with contralateral vascularized fibula transplantation. Bone healing was evaluated by regular X-ray examination. At the end of the follow-up, lower limb length, knee flexion, and extension function were evaluated. Patients were followed up for 24 to 36 months. Average bone-healing time was 5.2 months (3-8 months). All patients achieved bone healing with no tumor recurrence and no distant metastasis and all patients survived. The lengths of both lower limbs were equal in 2 cases, with shortening by ≤1 cm in 1 case and shortening by 2 cm in 1 case. Knee flexion was >90° in 4 cases and between 50 and 60° in 1 case. The Muscle and Skeletal Tumor Society score was 24.2 (range 20-26). Inactivation of autogenous bone with the epiphysis preserved by liquid nitrogen combined with vascularized fibula reconstruction for periarticular osteosarcoma of the knee in children is safe and effective. This technique supports bone healing. Postoperative limb length and function, and short-term effects were satisfactory.

Safety and efficacy of Pucotenlimab (HX008) - a humanized immunoglobulin G4 monoclonal antibody in patients with locally advanced or metastatic melanoma: a single-arm, multicenter, phase II study.

BACKGROUND: Pucotenlimab is a novel recombinant humanized anti-PD-1 (Programmed death-1) monoclonal antibody, which belongs to the human IgG4/kappa subtype, and can selectively block the binding of PD-1 with its ligands PD-L1 and PD-L2. METHODS: In this phase 2 trial, patients with locally advanced or metastatic melanoma who had failed conventional treatment (chemotherapy, targeted therapy, interferon, IL-2, et al.) were recruited. The patients were administrated with Pucotenlimab of 3 mg/kg every 3 weeks until disease progression, intolerable toxicity, or treatment discontinuation for any other reasons. The primary endpoint was the overall response rate (ORR). The secondary endpoints were disease control rate (DCR), duration of response (DOR), progression-free survival (PFS), overall survival (OS), and toxicity. RESULTS: One-hundred and nineteen patients were enrolled and followed up for 19.32 (ranging from 15.901 to 24.608) months by the cutoff date of July 30th, 2021. The ORR was 20.17% (24/119, 95% CI, 13.370%-28.506%) based on both independent review committee (IRC) and the investigator's assessment per RECIST v1.1. The median PFS were 2.89 (95% CI, 2.037-4.074) months and 2.46 (95% CI, 2.004-4.008) months based on IRC and investigator's assessment, respectively, per RECIST v1.1. The median OS was 16.59 (95% CI, 13.963-26.973) months. Treatment-related adverse events (TRAEs) occurred in 77.3% (92/119) of the patients. The incidence of Grade ≥ 3 TRAEs was 15.1% (18/119). In addition, none of the patients died because of TRAEs. As for biomarker analysis, Eotaxin (CCL11) and MCP-1 (CCL2) were related to treatment response, while TNF-α and VEGF were related to treatment failure. CONCLUSIONS: Pucotenlimab as a ≥ 2nd line therapy showed promising efficacy and tolerable toxicity for patients with locally advanced or metastatic melanoma. TRIAL REGISTRATION: Clinicaltrials.gov Identifier: NCT04749485 (registered retrospectively on 11/02/2021).

A Comprehensive Understanding of the Genomic Bone Tumor Landscape: A Multicenter Prospective Study.

Complexity and heterogeneity increases the difficulty of diagnosis and treatment of bone tumors. We aimed to identify the mutational characterization and potential biomarkers of bone tumors. In this study, a total of 357 bone tumor patients were recruited and the next generation sequencing (NGS)-based YuanSu450 panel, that includes both DNA and RNA sequencing, was performed for genomic alteration identification. The most common mutated genes in bone tumors included TP53, NCOR1, VEGFA, RB1, CCND3, CDKN2A, GID4, CCNE1, TERT, and MAP2K4. The amplification of genes such as NCOR1, VEGFA, and CCND3 mainly occurred in osteosarcoma. Germline mutation analysis reveal a high frequency of HRD related mutations (46.4%, 13/28) in this cohort. With the assistance of RNA sequencing, 16.8% (19/113) gene fusions were independently detected in 20% (16/79) of patients. Nearly 34.2% of patients harbored actionable targeted mutations, of which the most common mutation is CDKN2A deletion. The different mutational characterizations between juvenile patients and adult patients indicated the potential effect of age in bone tumor treatment. According to the genomic alterations, the diagnosis of 26 (7.28%) bone tumors were corrected. The most easily misdiagnosed bone tumor included malignant giant cell tumors of bone (2.8%, 10/357) and fibrous dysplasia of bone (1.7%, 6/357). Meanwhile, we found that the mutations of MUC16 may be a potential biomarker for the diagnosis of mesenchymal chondrosarcomas. Our results indicated that RNA sequencing effectively complements DNA sequencing and increased the detection rate of gene fusions, supporting that NGS technology can effectively assist the diagnosis of bone tumors.

Resection and reconstruction of huge tumors in the chest wall.

OBJECTIVE: To evaluate the experience and effects of resection and reconstruction of 4 cases of huge tumors in the chest wall. METHODS: The clinical data of 4 patients with huge tumors in the chest wall from July 2015 to January 2020 were collected and analyzed. There were 2 males and 2 females.Chondrosarcoma was diagnosed in 2 cases, giant cell tumor was diagnosed in 1 case,and metastasis from breast cancer was diagnosed in 1 case.All patients underwent extensive tumor resection and had thoracic exposure after tumor resection.Two patients underwent reconstruction with mesh and titanium mesh, and the incision was closed directly.The third patient underwent reconstruction with mesh and latissimus dorsi flap,and the fourth patient underwent reconstruction with mesh,titanium mesh and latissimus dorsi flap. RESULT: One patient had incision infection after operation,which resolved after debridement.All patients were followed up for 2-6 years, no tumor recurrence or metastasis was noted during follow-up.None of patients had abnormal breathing, dyspnea or other physical discomfort. CONCLUSION: It is difficult to resect the huge tumors in the chest wall,and it is more reasonable and safer to choose a reconstruction method using mesh and titanium mesh.The latissimus dorsi flap can achieve good results in repairing soft tissue defects.Close perioperative management and multidisciplinary team discussions can help to achieve better curative effects.

METTL3 stabilizes HDAC5 mRNA in an m6A-dependent manner to facilitate malignant proliferation of osteosarcoma cells.

Osteosarcoma (OS) is a prevalent primary bone sarcoma. Methyltransferase-like 3 (METTL3) is dysregulated in human malignancies. This study explored the mechanism of METTL3 in OS cell proliferation. Our results demonstrated that METTL3 was highly expressed in OS, and correlated with the tumor size, clinical stage, and distant metastasis of OS patients. Higher METTL3 expression indicated poorer prognosis. METTL3 silencing inhibited the malignant proliferation of OS cells, while METTL3 overexpression led to an opposite trend. METTL3 upregulated histone deacetylase 5 (HDAC5) expression in OS cells by increasing the m6A level. HDAC5 reduced the enrichment of H3K9/K14ac on miR-142 promoter, thus suppressing miR-142-5p expression and upregulating armadillo-repeat-containing 8 (ARMC8) level. HDAC5 overexpression or miR-142-5p silencing attenuated the inhibitory effect of METTL3 silencing on OS cell proliferation. Xenograft tumor experiment in nude mice confirmed that METTL3 silencing repressed OS cell proliferation in vivo via the HDAC5/miR-142-5p/ARMC8 axis. Collectively, METTL3-mediated m6A modification facilitated OS cell proliferation via the HDAC5/miR-142-5p/ARMC8 axis.

Clinical characteristics and prognosis of acral lentiginous melanoma: a single-center series of 211 cases in China.

BACKGROUND: Acral lentiginous melanoma (ALM) is common in China with poor prognosis. However, there are only a few studies of ALM in the Asian population. We aimed to summarize and analyze the clinical characteristics, treatment strategy, treatment effect, and prognostic factors of ALM in a Chinese population. METHODS: We included a total of 249 ALM patients (211 with follow-up data) from a single institution. Demographic, laboratory data, treatment strategy, and prognosis were analyzed. RESULTS: The ratio of male and female was 1.3 ∶ 1.0. The median age was 58 years old. The majority of patients (70.3%) had lesions on the sole. Trauma history and irritation were associated with lesion size increase in some patients. The prognosis of patients in stage II-III undergoing standard operation was significantly better compared with those without surgical treatment. Patients who did not receive postoperative adjuvant treatment had shorter time to distant metastasis. In multivariable analysis, distant metastasis, duration of disease, LDH level, and Ki67 index were independently associated with survival. CONCLUSIONS: Prognosis for ALM patients was poor in our study. Distant metastasis, duration of disease, LDH level, and Ki67 index were independently associated with prognosis.

Feasibility Analysis and Clinical Applicability of a Modified Type V Resection Method for Malignant Bone Tumors of the Proximal Humerus.

Objective: The purpose of this study was to explore the feasibility and clinical applicability of a modified type V resection method for malignant bone tumors of the proximal humerus. Methods: The relevant anatomic MRI data from 30 normal adult shoulder joints were measured to analyze the feasibility of the modified type V resection method for malignant bone tumors of the proximal humerus. Sixteen patients with malignant bone tumors of the proximal humerus were treated with modified radical resection between March 2012 and April 2017. Recurrence of tumor was evaluated after surgery, and shoulder function was assessed according to the Enneking skeletal muscle tumor function scoring system. Results: Radiographic results showed that the modified type V resection method was feasible, and within the allowable range of the maximum longitudinal diameter (<29.8 mm) and depth (<4 mm). Surgery was successfully completed in all 16 cases, and pathological examination suggested that the purposes for radical resection had been achieved. All patients were followed up over 3-49 months (mean, 15.6 months). One patient had local recurrence at 12 months after surgery, and we performed upper limb amputation. The remaining 15 patients had good prosthesis survival. At the final follow-up, shoulder joint function had recovered compared with preoperative levels, with a mean Enneking score of 25.8 points (range, 24-27 points). Conclusion: Modified type V resection may be feasible for treating tumors of the proximal humerus, maintaining good early shoulder function.

[FEASIBILITY AND SHORT-TERM EFFECTIVENESS ANALYSIS OF MODIFIED RADICAL RESECTION AND RECONSTRUCTION SURGERY FOR MALIGNANT PROXIMAL HUMERUS TUMOR].

OBJECTIVE: To explore the feasibility and short-term effectiveness of the modified radical resection and reconstruction in the treatment of malignant proximal humerus tumor. METHODS: The relevant anatomic data from 30 normal adult shoulder joint MRI were measured to analyze the feasibility of modified radical resection and reconstruction surgery in the treatment of malignant proximal humerus tumor. Five patients with malignant proximal humerus tumor were treated by using the modified radical resection and reconstruction surgery between March 2012 and January 2016. There were 1 male and 4 females, aged from 9 to 69 years (median, 46 years). There were 4 cases of osteosarcoma (Enneking IIA in 2 cases and Enneking IIB in 2 cases) and 1 case of metastatic carcinoma (moderately differentiated adenocarcinoma). The disease duration was 7 to 12 months (mean, 9 months). Recurrence of tumor was observed after operation, and the shoulder function was assessed according to Enneking skeletal muscle tumor function scoring system. RESULTS: Radiographic results showed that modified radical resection and reconstruction surgery was feasible, which was in allowable range of the maximum longitudinal diameter (<29.8 mm) and depth (<4 mm). The operation was successfully completed in all 5 cases, and pathological examination suggested that purposes of radical resection had achieved. All patients were followed up 3 to 49 months (mean, 15.6 months). One patient had local recurrence at 12 months after operation, and a shoulder joint amputation was performed; the other 4 patients had good prosthesis survival. At last follow-up, the function of the shoulder joint was obviously recovered when compared with preoperative function; Enneking's skeletal muscle tumor function score was 25.8 points (range, 24 to 27 points). CONCLUSIONS: Modified radical resection and reconstruction surgery is feasible for the treatment of proximal humerus tumor, and it can maintain a good early shoulder function.

Classification and Tie2 mutations in spinal and soft tissue vascular anomalies.

Vascular anomalies included hemangiomas and vascular malformations (VMs). VMs are mediated by mutations in the endothelial cell-specific receptor tyrosine kinase Tie2 (TEK),which is essential for angiogenesis and vascular stabilization. We identified five types of Tie2 mutations in 80 patients with soft tissue or spinal VMs by PCR including the previously detected missense mutations 2690A>G (Y897C), 2740C>T (L914F), 2743C>T (R915C), and two nonsense mutations 2763G>A, 2688C>T, we identified Tie2 mutation in primary spinal VMs for the first time. Tie2 mutations were found to be absent in 33 patients with hemangiomas and DNA samples of VMs. In addition, we showed that Tie2 mRNA expression in spinal VMs was similar to soft tissue VMs, but obviously lower than infant hemangiomas (P<0.01). This study provides new insights into spinal VMs, the association of Tie2 and vascular anomalies needs to be further discussed.