PRRX1-TOP2A interaction is a malignancy-promoting factor in human malignant peripheral nerve sheath tumours.

BACKGROUND: Paired related-homeobox 1 (PRRX1) is a transcription factor in the regulation of developmental morphogenetic processes. There is growing evidence that PRRX1 is highly expressed in certain cancers and is critically involved in human survival prognosis. However, the molecular mechanism of PRRX1 in cancer malignancy remains to be elucidated. METHODS: PRRX1 expression in human Malignant peripheral nerve sheath tumours (MPNSTs) samples was detected immunohistochemically to evaluate survival prognosis. MPNST models with PRRX1 gene knockdown or overexpression were constructed in vitro and the phenotype of MPNST cells was evaluated. Bioinformatics analysis combined with co-immunoprecipitation, mass spectrometry, RNA-seq and structural prediction were used to identify proteins interacting with PRRX1. RESULTS: High expression of PRRX1 was associated with a poor prognosis for MPNST. PRRX1 knockdown suppressed the tumorigenic potential. PRRX1 overexpressed in MPNSTs directly interacts with topoisomerase 2 A (TOP2A) to cooperatively promote epithelial-mesenchymal transition and increase expression of tumour malignancy-related gene sets including mTORC1, KRAS and SRC signalling pathways. Etoposide, a TOP2A inhibitor used in the treatment of MPNST, may exhibit one of its anticancer effects by inhibiting the PRRX1-TOP2A interaction. CONCLUSION: Targeting the PRRX1-TOP2A interaction in malignant tumours with high PRRX1 expression might provide a novel tumour-selective therapeutic strategy.

Soft-tissue sarcoma in adolescents and young adults.

Soft-tissue sarcoma is a rare cancer that accounts for approximately 1% of all malignant tumors. Although they occur in various age groups, soft-tissue sarcomas account for 8% of all malignant tumors developing in adolescents and young adults, suggesting that they are not rare in this age group. This study aimed to evaluate the clinical and pathological characteristics of soft-tissue sarcoma in adolescents and young adults. According to the Bone and Soft-Tissue Tumor Registry in Japan, myxoid liposarcoma is the most common type of soft-tissue sarcoma found in adolescents and young adults; alveolar soft part sarcoma, extraskeletal Ewing sarcoma, epithelioid sarcoma, clear cell sarcoma and synovial sarcoma occur predominantly in this age group among soft-tissue sarcomas. The analysis based on this registry demonstrated that age was not a prognostic factor for poor survival of soft-tissue sarcoma, although the prognosis in adolescents and young adults was better than that in older patients in the US and Scandinavia. Adolescent and young adult patients with soft-tissue sarcoma have age-specific problems, and a multidisciplinary approach to physical, psychological, and social issues is necessary to improve the management of these young patients both during and after treatment.

Medial Meniscus Posterior Root Repair Using a Modified Mason-Allen Suture Can Prevent the Progression of Cartilage Degeneration on the Loading Surface of the Medial Compartment: A Second-Look Arthroscopic Evaluation.

The treatment of medial meniscus posterior root tears (MMPRTs) has evolved to include a variety of repair strategies. This study investigated the location of the articular cartilage degeneration during second-look arthroscopy after transtibial pullout repair with a modified Mason-Allen suture using FasT-Fix (F-MMA) in 22 patients with MMPRTs. Second-look arthroscopy was performed approximately 1 year postoperatively to eval-uate the healing status of the medial meniscus (MM). Articular cartilage degeneration was assessed using the International Cartilage Repair Society grade at primary surgery and again at second-look arthroscopy. Articular surfaces of the medial/lateral femoral condyles, the medial/lateral tibial plateaus, the patella and the trochlea were divided into several subcompartments (MF 1-9, LF 1-9, MT 1-5, LT 1-5, P 1-9, T 1-3). Clinical evaluations used the Japanese Knee Injury and Osteoarthritis Outcome, Lysholm, and International Knee Documentation Committee scores. Second-look arthroscopic findings showed complete healing of the MM posterior root in all patients. Significant differences between pullout repair and second-look arthroscopy were observed for MF 2 and 4, LF 7, and P 7. All clinical outcomes were improved. Our results indicate that this technique improves clinical outcomes postoperatively and may prevent the progression of cartilage degenera-tion on the loading surface of the medial knee compartment.

Induction and expansion of human PRRX1+ limb-bud-like mesenchymal cells from pluripotent stem cells.

Current protocols for the differentiation of human pluripotent stem cells (hPSCs) into chondrocytes do not allow for the expansion of intermediate progenitors so as to prospectively assess their chondrogenic potential. Here we report a protocol that leverages PRRX1-tdTomato reporter hPSCs for the selective induction of expandable and ontogenetically defined PRRX1+ limb-bud-like mesenchymal cells under defined xeno-free conditions, and the prospective assessment of the cells' chondrogenic potential via the cell-surface markers CD90, CD140B and CD82. The cells, which proliferated stably and exhibited the potential to undergo chondrogenic differentiation, formed hyaline cartilaginous-like tissue commensurate to their PRRX1-expression levels. Moreover, we show that limb-bud-like mesenchymal cells derived from patient-derived induced hPSCs can be used to identify therapeutic candidates for type II collagenopathy and we developed a method to generate uniformly sized hyaline cartilaginous-like particles by plating the cells on culture dishes coated with spots of a zwitterionic polymer. PRRX1+ limb-bud-like mesenchymal cells could facilitate the mass production of chondrocytes and cartilaginous tissues for applications in drug screening and tissue engineering.

Adrenergic signaling promotes the expansion of cancer stem-like cells of malignant peripheral nerve sheath tumors.

Malignant peripheral nerve sheath tumor (MPNST), a highly malignant tumor that arises in peripheral nerve tissues, is known to be highly resistant to radiation and chemotherapy. Although there are several reports on genetic mutations and epigenetic changes that define the pathogenesis of MPNST, there is insufficient information regarding the microenvironment that contributes to the malignancy of MPNST. In the present study, we demonstrate that adrenaline increases the cancer stem cell population in MPNST. This effect is mediated by adrenaline stimulation of beta-2 adrenergic receptor (ADRB2), which activates the Hippo transducer, YAP/TAZ. Inhibition and RNAi experiments revealed that inhibition of ADRB2 attenuated the adrenaline-triggered activity of YAP/TAZ and subsequently attenuated MPNST cells stemness. Furthermore, ADRB2-YAP/TAZ axis was confirmed in the MPNST patients' specimens. The prognosis of patients with high levels of ADRB2 was found to be significantly worse. These data show that adrenaline exacerbates MPNST prognosis and may aid the development of new treatment strategies for MPNST.

Immunotherapy for sarcomas.

Sarcomas are a heterogeneous group of malignancies of mesenchymal origin; their molecular and genomic mechanisms differ with regard to histology. These characteristics lead to the presentation of varied immunological profiles based on the tumor microenvironment. Various immunotherapies are considered for the treatment of sarcoma. These treatments are performed either in isolation or in combination with other methods such as cytotoxic chemotherapy or the use of molecular target agents. Among these, two recently emerging immunotherapies include T-cell receptor gene therapy and immune checkpoint inhibitor therapy, which are expected to be effective for many types of sarcoma. A sarcoma with a disease-specific translocation and a limited number of mutations, such as synovial sarcoma, expresses high levels of self-antigens, like the New York esophageal squamous cell carcinoma 1, which has been targeted in T-cell receptor gene therapy. On the other hand, sarcomas with a greater number of mutations, such as undifferentiated pleomorphic sarcomas, myxofibrosarcoma and dedifferentiated liposarcomas, can be good candidates for immune checkpoint inhibitors. Among immune checkpoint inhibitor therapies, programmed cell death-1 blockade (nivolumab and pembrolizumab) and cytotoxic T-lymphocyte-associated antigen 4 blockade (ipilimumab) have been investigated most often in sarcoma. Although the sole use of immune checkpoint inhibitors provides limited efficacy, combined immunotherapy with immune checkpoint inhibitors or molecular target agents, especially antiangiogenic agents, has shown moderate results against some types of sarcoma, such as the alveolar soft part sarcoma. Several clinical trials utilizing immunotherapy, including T-cell receptor gene therapy and immune checkpoint inhibitors, in sarcomas are under progress. By clarifying the tumor microenvironment and biomarker-predictive capacity of immunotherapy in sarcomas, better clinical trials can be designed; this could lead to improved outcomes for immunotherapy in sarcoma.

PRRX1 promotes malignant properties in human osteosarcoma.

Paired related homeobox 1 (PRRX1) is a marker of limb bud mesenchymal cells, and deficiency of p53 or Rb in Prrx1-positive cells induces osteosarcoma in several mouse models. However, the regulatory roles of PRRX1 in human osteosarcoma have not been defined. In this study, we performed PRRX1 immunostaining on 35 human osteosarcoma specimens to assess the correlation between PRRX1 level and overall survival. In patients with osteosarcoma, the expression level of PRRX1 positively correlated with poor prognosis or the ratio of lung metastasis. Additionally, we found PRRX1 expression on in 143B cells, a human osteosarcoma line with a high metastatic capacity. Downregulation of PRRX1 not only suppressed proliferation and invasion but also increased the sensitivity to cisplatin and doxorubicin. When 143B cells were subcutaneously transplanted into nude mice, PRRX1 knockdown decreased tumor sizes and rates of lung metastasis. Interestingly, forskolin, a chemical compound identified by Connectivity Map analysis using RNA expression signatures during PRRX1 knockdown, decreased tumor proliferation and cell migration to the same degree as PRRX1 knockdown. These results demonstrate that PRRX1 promotes tumor malignancy in human osteosarcoma.

Steep posterior slope and shallow concave shape of the medial tibial plateau are risk factors for medial meniscus posterior root tears.

PURPOSE: Bone morphological factors are important for menisci. Their association with medial meniscus posterior root tears, however, has not yet been studied. This study aimed to compare sagittal medial tibial slope and medial tibial plateau depth between knees with and without medial meniscus posterior root tears. METHODS: Nine healthy volunteers, 24 patients who underwent anterior cruciate ligament reconstruction, and 36 patients who underwent medial meniscus posterior root pullout repair were included. Magnetic resonance imaging examinations were performed in the 10°-knee-flexed position. The medial tibial slope and medial tibial plateau depth were compared among the groups. RESULTS: In healthy volunteers, the anterior cruciate ligament reconstruction group, and the medial meniscus posterior root tear group, the medial tibial slopes were 3.5° ± 1.4°, 4.0° ± 1.9°, and 7.2° ± 1.9°, respectively, and the medial tibial plateau depths were 2.1 ± 0.7 mm, 2.2 ± 0.6 mm, and 1.2 ± 0.5 mm, respectively. Patients with medial meniscus posterior root tears had a significantly steep medial tibial slope and shallow medial tibial plateau concavity compared to those of healthy volunteers (P < 0.01) and the anterior cruciate ligament group (P < 0.01). In the multivariate logistic regression analysis, body mass index, medial tibial slope, and medial tibial plateau depth were significantly associated with medial meniscus posterior root tears. CONCLUSIONS: A steep posterior slope and a shallow concave shape of the medial tibial plateau are risk factors for medial meniscus posterior root tear. LEVEL OF EVIDENCE: Level III: Case-control study.

Medial meniscus posterior root repair prevents the progression of subchondral insufficiency fracture of the knee.

BACKGROUND: Medial meniscus posterior root tear (MMPRT) causes medial meniscus extrusion (MME) and leads to subchondral insufficiency fracture of the knee (SIFK). However, the progression of SIFK after MMPRT pullout repair remains unknown. This study aimed to investigate the progression of SIFK and compare clinical outcomes in patients with SIFK to those without SIFK after MMPRT pullout repair. We hypothesized that the progression of SIFK would be prevented by MMPRT pullout repair, and clinical outcomes would improve in all patients. METHODS: The SIFK grade (1-4) was evaluated using T2-fat suppression magnetic resonance imaging. Thirty-eight patients without SIFK (n = 22) and with low-grade SIFK (1 and 2; n = 16) who underwent MMPRT pullout repair were included. Preoperative factors, such as the duration from injury to the time of magnetic resonance imaging/surgery (weeks), femorotibial angle (degree), MME (mm), and clinical outcomes were evaluated, as well as the progression of SIFK. RESULTS: SIFK was identified in only 9 patients (grade 1) postoperatively. Significantly improved clinical outcomes were observed in all patients. Preoperative femorotibial angle, MME, and duration from injury to the time of magnetic resonance imaging/surgery were 177.1 ± 1.5°, 3.2 ± 1.6 mm, and 6.4 ± 7.0/10.1 ± 7.5 weeks, respectively. No significant difference in preoperative factors and clinical outcomes was observed between patients with SIFK and those without SIFK. CONCLUSIONS: MMPRT pullout repair prevented the progression of low-grade SIFK and improved clinical outcomes in all patients, although bone contusions (grade 1 SIFK) were not completely healed within 1 year. MMPRT pullout repair could be a good treatment option for optimizing clinical outcomes in patients with low-grade SIFK.

Transtibial pullout repair of medial meniscus posterior root tears: effects on the meniscus healing score and ICRS grade among patients with mild osteoarthritis of the knee.

PURPOSE: To assess the effects of transtibial pullout repair for medial meniscus posterior root tears (MMPRTs) among patients with early osteoarthritis of the knee as measured by the meniscus healing score and to determine whether the meniscus healing score correlates with the International Cartilage Repair Society (ICRS) grade progression. METHODS: Forty-seven patients with mild osteoarthritic knees (Kellgren-Lawrence grade ≤ 2 and varus alignment < 5°) who underwent transtibial pullout repair less than 3 months after MMPRT onset were assessed. The association between meniscus healing scores at 1 year postoperatively and cartilage damage of the medial compartment (medial femoral condyle [MFC] and medial tibial plateau [MTP]) were evaluated. The MFC was divided into six zones (A to F) and the MTP into two zones (G and H). The mean ICRS grade for each zone was compared between the primary surgery and second-look arthroscopy. The correlation between cartilage damage and meniscus healing status at the time of second-look arthroscopy in each zone was analysed. RESULTS: The mean time interval from injury to surgery was 63 days, and all clinical scores showed significant improvement. There were no significant differences in the extent of cartilage damage in areas B, C, E, or F (n.s.) for MFC or in areas G and H (n.s.) for MTP. The meniscus healing score and cartilage damage were correlated in the loading areas (B, C, E, and H; - 0.53, - 0.45, - 0.33, and - 0.38, respectively; p < 0.05). CONCLUSION: Transtibial pullout repair of MMPRTs among patients with mild osteoarthritic knees improved the clinical outcomes and showed a negative correlation between high meniscus healing scores and ICRS grades in the medial compartment loading area. This study suggests that early surgery should be undertaken for patients with mild osteoarthritic knee who develop MMPRTs. LEVEL OF EVIDENCE: Level IV.

Comparison of the clinical outcomes of transtibial pull-out repair for medial meniscus posterior root tear: Two simple stitches versus modified Mason-Allen suture.

BACKGROUND: Transtibial pullout repair of a medial meniscus posterior root tear (MMPRT) is a commonly used procedure, and several techniques have been reported. We hypothesised that pull-out repairs using two simple stitches (TSS) would have similar postoperative outcomes as those using the modified Mason-Allen suture with FasT-Fix (F-MMA). We aimed to investigate the clinical outcomes of these techniques, including the meniscal healing status and osteoarthritic change. METHODS: The data of 68 patients who underwent transtibial pull-out repair were retrospectively investigated. The patients were divided into two groups of 41 and 27 patients using F-MMA and TSS, respectively. The clinical outcomes were assessed preoperatively and at second-look arthroscopy (the mean period from surgery was one year) using the Knee injury and Osteoarthritis Outcome Score. The meniscal healing status, evaluated at second-look arthroscopy, was compared between the two groups. The cartilage damage was graded as per the classification of the International Cartilage Repair Society and compared at the primary surgery and second-look arthroscopy. RESULTS: Both groups showed significant improvement in each clinical score. No significant difference was seen in the clinical outcome scores and the meniscal healing status between the two groups at second-look arthroscopy. Moreover, no significant progression of cartilage damage was observed in both groups. Fourteen patients in the F-MMA group developed a complication of suture bar failures postoperatively; however, there were no complications in the TSS group. CONCLUSIONS: The TSS and F-MMA techniques showed favourable clinical outcomes and would be established as clinically useful techniques for the MMPRT treatment.

Bilateral Anterior Cruciate Ligament Tear Combined with Medial Meniscus Posterior Root Tear.

The case of an individual with a bilateral anterior cruciate ligament (ACL) tear combined with a medial meniscus (MM) posterior root tear is described. A 34-year-old Japanese man with bilateral ACL rupture that occurred > 10 years earlier was diagnosed with bilateral ACL tear combined with MM posterior root tear (MMPRT). We performed a transtibial pullout repair of the MMPRT with ACL reconstruction. The tibial tunnels for the MM posterior root repair and ACL reconstruction were created separately. Postoperatively, a good clinical outcome and meniscal healing were obtained. Our surgical technique may thus contribute to anatomical MM posterior root repair and ACL reconstruction.

Posttraumatic cartilage degradation progresses following anterior cruciate ligament reconstruction: A second-look arthroscopic evaluation.

BACKGROUND: Several studies have demonstrated that posttraumatic knee osteoarthritis progresses even after anterior cruciate ligament reconstruction. Few reports described zone-specific cartilaginous damages after anterior cruciate ligament reconstruction. This study aimed to compare the status of articular cartilage at anterior cruciate ligament reconstruction with that at second-look arthroscopy. METHODS: This study included 20 patients (20 knees, 10 males and 10 females, mean age 22.4 years, Body mass index 24.4 kg/m2) that underwent arthroscopic anatomic double-bundle anterior cruciate ligament reconstruction and second-look arthroscopy. Mean periods from injury to reconstruction and from reconstruction to second-look arthroscopy were 3.4 and 15.3 months, respectively. Cartilage lesions were evaluated arthroscopically in the 6 articular surfaces and 40 articular subcompartments independently, and these features were graded with the International Cartilage Repair Society articular cartilage injury classification; comparisons were made between the grades at reconstruction and at second-look arthroscopy. Furthermore, clinical outcomes were assessed at reconstruction and at second-look arthroscopy, using the Lysholm knee score, Tegner activity scale, International Knee Documentation Committee score, Knee injury and Osteoarthritis Outcome Score, side-to-side difference of the KT-2000 arthrometer, and pivot shift test. RESULTS: Each compartment showed a deteriorated condition at second-look arthroscopy compared with the pre-reconstruction period. A significant worsening of the articular cartilage was noted in all compartments except the lateral tibial plateau and was also observed in the central region of the medial femoral condyle and trochlea after reconstruction. However, each clinical outcome was significantly improved postoperatively. CONCLUSIONS: Good cartilage conditions were restored in most subcompartments at second-look arthroscopy. Furthermore, posttraumatic osteoarthritic changes in the patellofemoral and medial compartments progressed even in the early postoperative period, although good knee stability and clinical outcomes were obtained. Care is necessary regarding the progression of osteoarthritis and the appearance of knee symptoms in patients undergoing anterior cruciate ligament reconstruction.

Pullout repair using modified Mason-Allen suture induces better meniscal healing and superior clinical outcomes: A comparison between two surgical methods.

BACKGROUND: Pullout repairs of medial meniscus posterior root tears (MMPRTs) have many surgical options. However, there has been no reliable clinical study conducted to compare the superiority of each pullout repair technique. The current study hypothesized that pullout repairs using a modified Mason-Allen suture with FasT-Fix (F-MMA) would have several advantages in postoperative clinical outcomes and meniscal healing compared with single FasT-Fix. The aim of this study was to investigate the clinical usefulness of these two techniques in treating MMPRTs. METHODS: Thirty-eight patients who had complete MMPRTs were included. All patients underwent transtibial pullout repairs. To compare the clinical usefulness between pullout repairs using single FasT-Fix and F-MMA techniques, patients were divided into two groups. Second-look arthroscopic evaluations of meniscal healing were performed at one year postoperatively. Clinical outcomes were assessed using: Lysholm and visual analogue scale (VAS) pain scores, and Knee Injury and Osteoarthritis Outcome Score (KOOS). RESULTS: Single FasT-Fix and F-MMA pullout repairs improved clinical outcomes in patients with MMPRTs. At second-look arthroscopy, VAS pain, KOOS pain, and arthroscopic meniscal healing scores following F-MMA pullout repairs were superior to those after single FasT-Fix pullout repairs. CONCLUSIONS: This study demonstrated that F-MMA suture configuration obtained better meniscal healing and superior clinical outcomes compared with single FasT-Fix repairs in patients with MMPRTs. These results suggest that the F-MMA pullout repair may possibly reduce knee pain in arthroscopic treatments of MMPRTs.

Combined use of bFGF and GDF-5 enhances the healing of medial collateral ligament injury.

Basic fibroblast growth factor (bFGF) and growth and differentiation factor (GDF)-5 stimulate the healing of medial collateral ligament (MCL) injury. However, the effect of isolated and combined use of bFGF/GDF-5 remains still unclear. We investigated cellular proliferation and migration responding to bFGF/GDF-5 using rabbit MCL fibroblasts. Rabbit MCL injury was treated by bFGF and/or GDF-5 with peptide hydrogels. Gene expression and deposition of collagens in healing tissues were evaluated. bFGF/GDF-5 treatment additively enhanced cell proliferation and migration. bFGF/GDF-5 hydrogels stimulated Col1a1 expression without increasing Col3a1 expression. Combined use of bFGF/GDF-5 stimulated type I collagen deposition and the reorganization of fiber alignment, and induced better morphology of fibroblasts in healing MCLs. Our study indicates that combined use of bFGF/GDF-5 might enhance MCL healing by increasing proliferation and migration of MCL fibroblasts, and by regulating collagen synthesis and connective fiber alignment.