From the identification of actionable molecular targets to the generation of faithful neuroblastoma patient-derived preclinical models.

BACKGROUND: Neuroblastoma (NB) represents the most frequent and aggressive form of extracranial solid tumor of infants. Although the overall survival of patients with NB has improved in the last years, more than 50% of high-risk patients still undergo a relapse. Thus, in the era of precision/personalized medicine, the need for high-risk NB patient-specific therapies is urgent. METHODS: Within the PeRsonalizEd Medicine (PREME) program, patient-derived NB tumors and bone marrow (BM)-infiltrating NB cells, derived from either iliac crests or tumor bone lesions, underwent to histological and to flow cytometry immunophenotyping, respectively. BM samples containing a NB cells infiltration from 1 to 50 percent, underwent to a subsequent NB cells enrichment using immune-magnetic manipulation. Then, NB samples were used for the identification of actionable targets and for the generation of 3D/tumor-spheres and Patient-Derived Xenografts (PDX) and Cell PDX (CPDX) preclinical models. RESULTS: Eighty-four percent of NB-patients showed potentially therapeutically targetable somatic alterations (including point mutations, copy number variations and mRNA over-expression). Sixty-six percent of samples showed alterations, graded as "very high priority", that are validated to be directly targetable by an approved drug or an investigational agent. A molecular targeted therapy was applied for four patients, while a genetic counseling was suggested to two patients having one pathogenic germline variant in known cancer predisposition genes. Out of eleven samples implanted in mice, five gave rise to (C)PDX, all preserved in a local PDX Bio-bank. Interestingly, comparing all molecular alterations and histological and immunophenotypic features among the original patient's tumors and PDX/CPDX up to second generation, a high grade of similarity was observed. Notably, also 3D models conserved immunophenotypic features and molecular alterations of the original tumors. CONCLUSIONS: PREME confirms the possibility of identifying targetable genomic alterations in NB, indeed, a molecular targeted therapy was applied to four NB patients. PREME paves the way to the creation of clinically relevant repositories of faithful patient-derived (C)PDX and 3D models, on which testing precision, NB standard-of-care and experimental medicines.

Alpha-1 Adrenergic Antagonists Sensitize Neuroblastoma to Therapeutic Differentiation.

Neuroblastoma (NB) is an aggressive childhood tumor, with high-risk cases having a 5-year overall survival probability of approximately 50%. The multimodal therapeutic approach for NB includes treatment with the retinoid isotretinoin (13-cis retinoic acid; 13cRA), which is used in the post-consolidation phase as an antiproliferation and prodifferentiation agent to minimize residual disease and prevent relapse. Through small-molecule screening, we identified isorhamnetin (ISR) as a synergistic compound with 13cRA in inhibiting up to 80% of NB cell viability. The synergistic effect was accompanied by a marked increase in the expression of the adrenergic receptor α1B (ADRA1B) gene. Genetic knockout of ADRA1B or its specific blockade using α1/α1B adrenergic antagonists led to selective sensitization of MYCN-amplified NB cells to cell viability reduction and neural differentiation induced by 13cRA, thus mimicking ISR activity. Administration of doxazosin, a safe α1-antagonist used in pediatric patients, in combination with 13cRA in NB xenografted mice exerted marked control of tumor growth, whereas each drug alone was ineffective. Overall, this study identified the α1B adrenergic receptor as a pharmacologic target in NB, supporting the evaluation of adding α1-antagonists to the post-consolidation therapy of NB to more efficiently control residual disease. SIGNIFICANCE: Targeting α-adrenergic receptors synergizes with isotretinoin to suppress growth and to promote differentiation of neuroblastoma, revealing a combinatorial approach for more effective management of the disease and prevention of relapse.

Clinical Comparison Between Shortening Osteotomy of the Proximal Phalanx Neck and Arthrodesis in Hammer Toe Surgery at Mid-TermFollow-Up.

The treatment of hammer toe has been advocated for years, and many procedures have been proposed with skeletal and soft tissue intervention. The purpose of the present study was to compare arthrodesis of the proximal interphalangeal joint with shortening osteotomy of the proximal phalanx neck. In most cases, both procedures have been associated with elongation of the extensor apparatus, capsulotomy of the metatarsophalangeal joint, and stabilization with a Kirschner wire. To experiment with a technique that respects the anatomy and joint function, we used a distal subtraction osteotomy of the proximal phalanx neck. We compared a series of 78 patients, divided in to 2 groups: 38 (48.7%) treated with arthrodesis and 40 (51.3%) with shortening osteotomy. Patients were aged 22 to 78 years, with a mean final follow-up period of 56.6 (range 24 to 96) months. For clinical evaluation, we used the American Orthopaedic Foot and Ankle Society score, Foot and Ankle Outcome Score, and a subjective rating scale. The results were comparable between the 2 techniques; however, we report faster functional recovery in the group treated with shortening osteotomy (p < .0001), with an adjunctive advantage of preserving the integrity of the proximal interphalangeal joint. Thus, according to our results, this technique is comparable to arthrodesis.

Multidirectional Shoulder Instability: Arthroscopic Labral Augmentation.

Capsulolabral augmentation is one of the most used arthroscopic techniques to address multidirectional instability of the shoulder. Given the thin and weak capsule seen in the affected patients, reconstruction in this subset of patients can be particularly challenging. This arthroscopic technique aims to reduce the capsular volume and deepen the glenoid socket through the creation of a particularly voluminous "bumper" along the glenoid bone. Increasing the depth of the glenoid facilitates a concavity-compression stabilizing effect and, therefore, shoulder stability, especially midrange stability. This technique aims to augment the bump of the standard capsulolabral reconstruction by using a resorbable surgical mesh derived from porcine skin.

Arthroscopic treatment of the atraumatic shoulder instability: a case series with two-year follow-up evaluation.

BACKGROUND: The purpose of this work is to evaluate the results of arthroscopic capsulolabroplasty in patients affected by atraumatic shoulder instability (ASI). METHODS: A retrospective review was performed of 10 patients (7 women and 3 men) who underwent arthroscopic treatment of symptomatic ASI. Mean age at evaluation was 27.9 (19-35) years and the mean follow-up was 23.3 (12-37) months. We evaluated recurrence rate, range of movement, apprehension and relocation tests, hyperlaxity, and sport activity. The ASES score, the Rowe score, the Simple Shoulder Test (SST) and Visual Analogue Scale (VAS) were also used as outcomes measure. RESULTS: None of the patients experienced episodes of dislocation or subluxation after surgery. The apprehension and relocation tests produced positive results in 2 patients. Six out of 10 patients reported apprehension with the arm in specific positions. The ASES mean score was 93.4 (55-100); the Rowe mean score was 85.5 (70-100); the SST mean score was 9.1 (5.8-10). On average, external rotation is reduced by 10° in adduction, and by 8° in abduction in 6 out of 10 patients; internal rotation is reduced on average by 6.6° in abduction with the arm abducted, and was overall limited in 6 out of 10 patients. CONCLUSIONS: Arthroscopic capsulolabroplasty ensures excellent results in patients showing atraumatic shoulder instability in terms of recurrence. Still, an underlying insecurity persists and the risk of residual stiffness is tangible. LEVEL OF EVIDENCE: V.

No-holes transpectoral tenodesis technique vs tenotomy of the long head of the biceps brachii.

BACKGROUND: There is no univocal consensus regarding Long Head of the Biceps (LHB) best treatment between tenotomy and tenodesis. There is no consensus regarding the best location to perform the tenodesis. The LHB tenodesis performed by the proximal tendon excision as first step can miss the proper tension to the muscle belly. Fixations proximal to the pectoralis major can lead to groove pain. This study aims to test the efficacy of a new LHB tenodesis technique by comparing its results with the tenotomy. METHODS: We retrospectively evaluated patients who underwent surgery between May 2014 and May 2015. The mean follow up was 14.7 months. Sixteen patients underwent mini-open tenodesis to the Pectoralis Major tendon by the use of a resorbable suture (TD group); sixteen underwent tenotomy (TT group). The mean age of the TD group was 54 years; the mean age of the TT group was 56 years. We evaluated pain, subjective perception of the patient of possible aesthetic and strength differences between the two biceps, "Popeye sign", and tests to stimulate the LHB. We administered three evaluation questionnaires: the ASES score, the SPADI score, and the SST. RESULTS: 32 consecutive patients were evaluated. The clinical scores did not record statistically significant differences: the mean ASES score was 92.9 (TD) and 90.8 (TT); the mean SPADI score was 92.5 (TD), and 89.7 (TT); the mean SST was 8.9 (TD), and 8.4 (TT). Compared to the TD group, in the TT group we registered with greater frequency the "Popeye sign" with a P value < 0.05 (9 cases vs 1), and spasms in the biceps muscle belly (5 cases vs 1). All other signs or symptoms evaluated were more frequent in the TT group, except the strength difference perceived by the patient (3 patients in the TT group, and 2 in the TD group). No complications were recorded. CONCLUSIONS: This new Long Head of the Biceps (LHB) tenodesis technique is valuable and reliable, and provided better results than tenotomy. LEVEL OF EVIDENCE: IV.

ACL Reconstruction: Choosing the Graft.

Rupture of the anterior cruciate ligament is one of the most common ligament injuries in sports traumatology. The need for surgical anterior cruciate ligament reconstruction is justified by its anatomical characteristics. Key considerations when choosing a graft include the potential for bone integration and the risk of failure. Bone sclerosis around the tunnel affects the integration of the graft. For this reason, one aspect upon which orthopedic surgeons should focus is the biology of the bone-graft interface. Although the BPTB graft is still used, hamstrings and synthetic grafts have become increasingly widespread and popular over the years. An allograft certainly requires more long-term follow-up to validate its use in response to functional, clinical and biological requirements.