Unveiling the role of osteosarcoma-derived secretome in premetastatic lung remodelling.

BACKGROUND: Lung metastasis is the most adverse clinical factor and remains the leading cause of osteosarcoma-related death. Deciphering the mechanisms driving metastatic spread is crucial for finding open therapeutic windows for successful organ-specific interventions that may halt or prevent lung metastasis. METHODS: We employed a mouse premetastatic lung-based multi-omics integrative approach combined with clinical features to uncover the specific changes that precede lung metastasis formation and identify novel molecular targets and biomarker of clinical utility that enable the design of novel therapeutic strategies. RESULTS: We found that osteosarcoma-bearing mice or those preconditioned with the osteosarcoma cell secretome harbour profound lung structural alterations with airway damage, inflammation, neutrophil infiltration, and extracellular matrix remodelling with increased deposition of fibronectin and collagens by resident stromal activated fibroblasts, favouring the adhesion of disseminated tumour cells. Systemic-induced microenvironmental changes, supported by transcriptomic and histological data, promoted and accelerated lung metastasis formation. Comparative proteome profiling of the cell secretome and mouse plasma identified a large number of proteins involved in extracellular-matrix organization, cell-matrix adhesion, neutrophil degranulation, and cytokine-mediated signalling, consistent with the observed lung microenvironmental changes. Moreover, we identified EFEMP1, an extracellular matrix glycoprotein exclusively secreted by metastatic cells, in the plasma of mice bearing a primary tumour and in biopsy specimens from osteosarcoma patients with poorer overall survival. Depletion of EFEMP1 from the secretome prevents the formation of lung metastasis. CONCLUSIONS: Integration of our data uncovers neutrophil infiltration and the functional contribution of stromal-activated fibroblasts in ECM remodelling for tumour cell attachment as early pro-metastatic events, which may hold therapeutic potential in preventing or slowing the metastatic spread. Moreover, we identified EFEMP1, a secreted glycoprotein, as a metastatic driver and a potential candidate prognostic biomarker for lung metastasis in osteosarcoma patients. Osteosarcoma-derived secreted factors systemically reprogrammed the lung microenvironment and fostered a growth-permissive niche for incoming disseminated cells to survive and outgrow into overt metastasis. Daily administration of osteosarcoma cell secretome mimics the systemic release of tumour-secreted factors of a growing tumour in mice during PMN formation; Transcriptomic and histological analysis of premetastatic lungs revealed inflammatory-induced stromal fibroblast activation, neutrophil infiltration, and ECM remodelling as early onset pro-metastatic events; Proteome profiling identified EFEMP1, an extracellular secreted glycoprotein, as a potential predictive biomarker for lung metastasis and poor prognosis in osteosarcoma patients. Osteosarcoma patients with EFEMP1 expressing biopsies have a poorer overall survival.

ANKLE ARTHRODESIS WITH INTRAMEDULLARY RETROGRADE NAIL FOR BONE TUMORS. PRELIMINARY RESULTS AND SURGICAL TECHNIQUE.

Objective: Present the preliminary results of a case series using the surgical ankle arthrodesis technique with an intramedullary retrograde nail for bone tumors. Methods: We present the preliminary data of 4 patients, 3 males and 1 female, with a mean age of 46,2 (range 32 to 58) years, with histology proven Giant Cell Tumour of bone in 3 and osteosarcoma in 1. The mean resection length of distal tibia was 11,75 (range 9 to 16) cm, and all the patients underwent reconstruction with a tibiotalocalcaneal arthrodesis with an intercalary allograft fixed by a retrograde intramedullary nail. Results: Oncological follow-up evolved without evidence of local recurrence or disease progression in all patients. After a mean time of 69.5 (range 32 to 98 months), patients had a mean MSTS12 functional score of 82.5% (range 75 to 90). All tibial arthrodesis and diaphyseal osteotomy sites were fused within 6 months with a return to activities without complications related to coverage skin or infection. Conclusion: No complications were recorded; all arthrodesis and diaphysial tibial osteotomy sites fused by 6 months, and the mean follow-up of those patients was 69,5 (range 32 to 988) months, with a mean functional MSTS score of 82,5% (range 75-90). Level of Evidence: IV; Retrospective Case Series.

Objetivo: Apresentar os resultados preliminares de uma série de casos utilizando a técnica cirúrgica de artrodese do tornozelo com haste intramedular retrógada para tumores ósseos. Métodos: Apresentamos os dados preliminares de quatro pacientes, três homens e uma mulher, com idade média de 46,2 (variação de 32 a 58) anos, com histologia comprovada de tumor de células gigantes em três e osteossarcoma em um. O comprimento médio de ressecção da tíbia distal foi de 11,75 (variação de 9 a 16) cm, e todos os pacientes foram submetidos à reconstrução com uma artrodese tibiotalocalcaneana com um aloenxerto intercalar fixado por uma haste intramedular retrógrada. Resultados: O acompanhamento oncológico evoluiu sem evidências de recidiva local ou progressão da doença, em todos os pacientes. Após um tempo médio de 69,5 (variação de 32 a 98 meses), os pacientes tiveram uma pontuação média funcional MSTS12 de 82,5% (variação de 75 a 90). Todos os locais de artrodese e osteotomia diafisária tibiais foram fundidos em 6 meses com retorno às atividades de vida diária sem complicações relacionadas à cobertura ou infecção. Conclusão: Não foram registradas complicações; todos os locais de artrodese e osteotomia diafisária da tíbia fundiram-se em 6 meses, e o acompanhamento médio desses pacientes foi de 69,5 (variação de 32 a 988) meses, com uma pontuação média funcional MSTS de 82,5% (variação de 75-90). Nível de Evidência IV; Série de Casos Retrospectivos.

Natural Killer T-like Cells: Immunobiology and Role in Disease.

CD56+ T cells are generally recognized as a distinct population of T cells and are categorized as NKT-like cells. Although our understanding of NKT-like cells is far from satisfactory, it has been shown that aging and a number of disease situations have impacted these cells. To construct an overview of what is currently known, we reviewed the literature on human NKT-like cells. NKT-like cells are highly differentiated T cells with "CD1d-independent" antigen recognition and MHC-unrestricted cell killing. The genesis of NKT-like cells is unclear; however, it is proposed that the acquisition of innate characteristics by T cells could represent a remodeling process leading to successful aging. Additionally, it has been shown that NKT-like cells may play a significant role in several pathological conditions, making it necessary to comprehend whether these cells might function as prognostic markers. The quantification and characterization of these cells might serve as a cutting-edge indicator of individual immune health. Additionally, exploring the mechanisms that can control their killing activity in different contexts may therefore result in innovative therapeutic alternatives in a wide range of disease settings.

Ankle arthrodesis with intramedullary retrograde nail for bone tumors. preliminary results and surgical technique / Artrodese do tornozelo com cavilha intramedular retrógrada para tumores ósseos. resultados preliminares e técnica cirúrgica

ABSTRACT Objective Present the preliminary results of a case series using the surgical ankle arthrodesis technique with an intramedullary retrograde nail for bone tumors. Methods We present the preliminary data of 4 patients, 3 males and 1 female, with a mean age of 46,2 (range 32 to 58) years, with histology proven Giant Cell Tumour of bone in 3 and osteosarcoma in 1. The mean resection length of distal tibia was 11,75 (range 9 to 16) cm, and all the patients underwent reconstruction with a tibiotalocalcaneal arthrodesis with an intercalary allograft fixed by a retrograde intramedullary nail. Results Oncological follow-up evolved without evidence of local recurrence or disease progression in all patients. After a mean time of 69.5 (range 32 to 98 months), patients had a mean MSTS12 functional score of 82.5% (range 75 to 90). All tibial arthrodesis and diaphyseal osteotomy sites were fused within 6 months with a return to activities without complications related to coverage skin or infection. Conclusion No complications were recorded; all arthrodesis and diaphysial tibial osteotomy sites fused by 6 months, and the mean follow-up of those patients was 69,5 (range 32 to 988) months, with a mean functional MSTS score of 82,5% (range 75-90). Level of Evidence: IV; Retrospective Case Series.

RESUMO Objetivo Apresentar os resultados preliminares de uma série de casos utilizando a técnica cirúrgica de artrodese do tornozelo com haste intramedular retrógada para tumores ósseos. Métodos Apresentamos os dados preliminares de quatro pacientes, três homens e uma mulher, com idade média de 46,2 (variação de 32 a 58) anos, com histologia comprovada de tumor de células gigantes em três e osteossarcoma em um. O comprimento médio de ressecção da tíbia distal foi de 11,75 (variação de 9 a 16) cm, e todos os pacientes foram submetidos à reconstrução com uma artrodese tibiotalocalcaneana com um aloenxerto intercalar fixado por uma haste intramedular retrógrada. Resultados O acompanhamento oncológico evoluiu sem evidências de recidiva local ou progressão da doença, em todos os pacientes. Após um tempo médio de 69,5 (variação de 32 a 98 meses), os pacientes tiveram uma pontuação média funcional MSTS12 de 82,5% (variação de 75 a 90). Todos os locais de artrodese e osteotomia diafisária tibiais foram fundidos em 6 meses com retorno às atividades de vida diária sem complicações relacionadas à cobertura ou infecção. Conclusão Não foram registradas complicações; todos os locais de artrodese e osteotomia diafisária da tíbia fundiram-se em 6 meses, e o acompanhamento médio desses pacientes foi de 69,5 (variação de 32 a 988) meses, com uma pontuação média funcional MSTS de 82,5% (variação de 75-90). Nível de Evidência IV; Série de Casos Retrospectivos.

Tumor-Infiltrating Lymphocytes and Cancer Markers in Osteosarcoma: Influence on Patient Survival.

Osteosarcoma (OST) is the most common type of high-grade primary bone tumor, which mainly affects young adults. The current standard of care for OST combines surgical resection with chemotherapy. The clinical outcomes and the current options to treat OST patients are unsatisfactory and novel treatment strategies are needed. The crosstalk between tumor cells and immune cells is essential to the OST microenvironment. Despite the efforts that have been made to address the importance of immune-related factors in OST, there is still a lot to understand. The purpose of the current study was to evaluate the tumor-infiltrating lymphocytes (TIL), the expression of proteins involved in tumor biology, and their impact on the clinical outcome of OST patients. We studied 93 samples of OST patients using immunohistochemistry and histomorphometry. We looked for the infiltration of CD3+, CD4+, CD8+, TIA1+ and CD20+ cells and for the expression of CD44 standard (CD44s) and variant 6 (CD44v6), CD95/Fas, Fas-L, p53 and p-glycoprotein. All the parameters were analyzed for the influence on the occurrence of death and metastasis, plus patient overall survival (OS) and progression-free survival (PFS). The effect of sex, age, tumor location (distal femur or proximal tibia) and the combination with neoadjuvant chemotherapy was also assessed. Our results suggest that the presence of tumor-infiltrating CD4+ cells provides protection to OST patients, and that CD8+ cells have a significant impact on the patient's overall survival (OS) and progression-free survival (PFS), which is more evident in male patients. In addition, a strong association between tumor-infiltrating CD4+ cells and the presence of CD44s expression in tumor samples was observed. Analysis of TIL and tumor markers related to tumor biology could be useful to stratify patients and monitor the response to therapy, as well as to assist with the development of immunotherapy strategies to improve the effects of cytotoxic TIL to eradicate the tumor cells.

The Role of Natural Killer Cells in Soft Tissue Sarcoma: Prospects for Immunotherapy.

Soft-tissue sarcomas (STS) represent about 80% of sarcomas, and are a heterogeneous group of rare and malignant tumors. STS arise from mesenchymal tissues and can grow into structures such as adipose tissue, muscles, nervous tissue and blood vessels. Morphological evaluation has been the standard model for the diagnosis of sarcomas, and even in samples with similar characteristics, they present a diversity in cytogenetic and genetic sequence alterations, which further increases the diversity of sarcomas. This variety is one of the main challenges for the classification and understanding of STS patterns, as well as for their respective treatments, which further decreases patient survival (<5 years). Despite some studies, little is known about the immunological profile of STS. As for the immunological profile of STS in relation to NK cells, there is also a shortage of studies. Observations made in solid tumors show that the infiltration of NK cells in tumors is associated with a good prognosis of the disease. Notwithstanding the scarcity of studies to characterize NK cells, their receptors, and ligands in STS, it is noteworthy that the progression of these malignancies is associated with altered NK phenotypes. Despite the scarcity of information on the function of NK cells, their phenotypes and their regulatory pathways in STS, the findings of this study support the additional need to explore NK cell-based immunotherapy in STS further. Some clinical trials, very tentatively, are already underway. STS clinical trials are still the basis for adoptive NK-cell and cytokine-based therapy.

Tumor and Peripheral Immune Status in Soft Tissue Sarcoma: Implications for Immunotherapy.

Soft Tissue Sarcomas (STS) are a heterogeneous and rare group of tumors. Immune cells, soluble factors, and immune checkpoints are key elements of the complex tumor microenvironment. Monitoring these elements could be used to predict the outcome of the disease, the response to therapy, and lead to the development of new immunotherapeutic approaches. Tumor-infiltrating B cells, Natural Killer (NK) cells, tumor-associated neutrophils (TANs), and dendritic cells (DCs) were associated with a better outcome. On the contrary, tumor-associated macrophages (TAMs) were correlated with a poor outcome. The evaluation of peripheral blood immunological status in STS could also be important and is still underexplored. The increased lymphocyte-to-monocyte ratio (LMR) and neutrophil-to-lymphocyte ratio (NLR), higher levels of monocytic myeloid-derived suppressor cells (M-MDSCs), and Tim-3 positive CD8 T cells appear to be negative prognostic markers. Meanwhile, NKG2D-positive CD8 T cells were correlated with a better outcome. Some soluble factors, such as cytokines, chemokines, growth factors, and immune checkpoints were associated with the prognosis. Similarly, the expression of immune-related genes in STS was also reviewed. Despite these efforts, only very little is known, and much research is still needed to clarify the role of the immune system in STS.

Dermatological Manifestations in the Intensive Care Unit: A Practical Approach.

Dermatological problems are not usually related to intensive medicine because they are considered to have a low impact on the evolution of critical patients. Despite this, dermatological manifestations (DMs) are relatively frequent in critically ill patients. In rare cases, DMs will be the main diagnosis and will require intensive treatment due to acute skin failure. In contrast, DMs can be a reflection of underlying systemic diseases, and their identification may be key to their diagnosis. On other occasions, DMs are lesions that appear in the evolution of critical patients and are due to factors derived from the stay or intensive treatment. Lastly, DMs can accompany patients and must be taken into account in the comprehensive pathology management. Several factors must be considered when addressing DMs: on the one hand, the moment of appearance, morphology, location, and associated treatment and, on the other hand, aetiopathogenesis and classification of the cutaneous lesion. DMs can be classified into 4 groups: life-threatening DMs (uncommon but compromise the patient's life); DMs associated with systemic diseases where skin lesions accompany the pathology that requires admission to the intensive care unit (ICU); DMs secondary to the management of the critical patient that considers the cutaneous manifestations that appear in the evolution mainly of infectious or allergic origin; and DMs previously present in the patient and unrelated to the critical process. This review provides a characterization of DMs in ICU patients to establish a better identification and classification and to understand their interrelation with critical illnesses.

Long-Term Response after 94 Cycles of Trabectedin in a Patient with Metastatic Leiomyosarcoma of the Lower Extremity.

Leiomyosarcomas of the lower extremity are extremely rare disorders and account for 10-15% of limb soft tissue sarcomas. These tumours have poor prognosis and even in early stages, patients persist at high risk for local and distant relapse; consequently, the treatment of advanced leiomyosarcoma of the lower extremity embodies a substantial defy. We present the case of a 73-year-old man diagnosed with metastatic lower extremity leiomyosarcoma of the hallux soft tissue, and with bone, lung and lymph node metastasis. After core needle biopsy confirmation of high-grade fusocellular sarcoma, the patient underwent surgery of the primary tumour and received anthracycline-based chemotherapy. However, after a 7-month progression-free survival period, a CT revealed lung disease progression. Sequentially, the patient was treated with trabectedin (Yondelis®) at a dose of 1.5 mg/m2 resulting in complete remission of the lung metastasis and stable disease of the remaining lesions after 26 months of treatment. Afterwards, the patient started on maintenance therapy with trabectedin, resulting in long-lasting stable disease, as he was able to receive 94 cycles with very acceptable quality of life. Finally, in March 2019, the patient died of community-acquired pneumonia without objective progression disease. This clinical case reports the first patient ever treated with 94 cycles of trabectedin. Our results additionally confirm that trabectedin wields relevant oncostatic benefits with a manageable safety profile and without cumulative toxicities. Trabectedin properties enable a maintenance long-term therapy (until disease progression or unbearable toxicity), with a high impact on survival and with a preserved quality of life.

Resurfaced allograft-prosthetic composite for proximal tibial reconstruction in children: intermediate-term results of an original technique.

BACKGROUND: Reconstruction of the proximal part of the tibia in children with megaprostheses or standard composite devices removes the distal femoral physis, and osteoarticular allografts have a high incidence of mechanical failure. We evaluated the results of an alternative original technique for proximal tibial reconstruction after tumor resection. METHODS: Nineteen children, ranging in age from six to sixteen years, who underwent surgical resection and reconstruction with a resurfaced allograft-prosthetic composite were retrospectively reviewed at a mean follow-up time of seventy-eight months. Eighteen children had high-grade sarcoma (fourteen had osteosarcoma and four had Ewing sarcoma), and one child had an aggressive giant cell tumor of the proximal part of the tibia. Surgical resection and its complications, disease-free survival of the patients, implant survival, and functional results according to the Musculoskeletal Tumor Society scoring system were analyzed at the time of the latest follow-up. RESULTS: In all patients, wide surgical margins were obtained and no local recurrence was observed. At the time of the latest follow-up, nine of the eighteen patients with a high-grade malignant tumor were continually disease-free, another six had had metastases but had no evidence of disease, and three had died of the disease. The patient with a giant cell tumor was continually disease-free after 168 months. The mean implant survival was sixty-eight months (range, six to 188 months). The major complications were one deep infection; two nonunions, which healed after surgical revision; and six allograft fractures. Limb-length discrepancy was the most frequent complication but, following contralateral epiphysiodesis, resulted in a mean discrepancy of 1.9 cm at the time of the latest follow-up. At the time of the latest follow-up, eight patients had a Musculoskeletal Tumor Society score deemed as excellent or good. CONCLUSIONS: In children and young teenagers with bone tumors of the proximal part of the tibia requiring an intra-articular resection, knee reconstruction with a resurfaced allograft-prosthetic composite may represent a valid alternative to conventional megaprostheses or to osteoarticular allograft.