TEG011 persistence averts extramedullary tumor growth without exerting off-target toxicity against healthy tissues in a humanized HLA-A*24:02 transgenic mice.

γδT cells play an important role in cancer immunosurveillance and are able to distinguish malignant cells from their healthy counterparts via their γδTCR. This characteristic makes γδT cells an attractive candidate for therapeutic application in cancer immunotherapy. Previously, we have identified a novel CD8α-dependent tumor-specific allo-HLA-A*24:02-restricted Vγ5Vδ1TCR with potential therapeutic value when used to engineer αßT cells from HLA-A*24:02 harboring individuals. αßT cells engineered to express this defined Vγ5Vδ1TCR (TEG011) have been suggested to recognize spatial changes in HLA-A*24:02 present selectively on tumor cells but not their healthy counterparts. However, in vivo efficacy and toxicity studies of TEG011 are still limited. Therefore, we extend the efficacy and toxicity studies as well as the dynamics of TEG011 in vivo in a humanized HLA-A*24:02 transgenic NSG (NSG-A24:02) mouse model to allow the preparation of a first-in-men clinical safety package for adoptive transfer of TEG011. Mice treated with TEG011 did not exhibit any graft-versus-host disease-like symptoms and extensive analysis of pathologic changes in NSG-A24:02 mice did not show any off-target toxicity of TEG011. However, loss of persistence of TEG011 in tumor-bearing mice was associated with the outgrowth of extramedullary tumor masses as also observed for mock-treated mice. In conclusion, TEG011 is well tolerated without harming HLA-A*24:02+ expressing healthy tissues, and TEG011 persistence seems to be crucial for long-term tumor control in vivo.

Selenium in the prevention and subsidiary therapy of cancer of soft tissues.

Selenium as an antioxidant has attracted attention because of its anticancer activity. This review presents a view on selenium and its compounds exerting influence against cancer in the soft tissues. The results reveal a significant strong association between a low selenium level in blood and a cancer risk. Seleno-supplementation is important in the prevention of metastatic cancer. These results help to elucidate the anticancer effect of selenium providing further evidence to exploit novel anticancer agents targeting selenium-containing organic compounds. Key words: selenium • cancer prevention • cancer treatment • soft tissues.

Radiation Therapy to Sites of Metastatic Disease as Part of Consolidation in High-Risk Neuroblastoma: Can Long-term Control Be Achieved?

PURPOSE: As part of consolidative therapy in high-risk neuroblastoma, modern protocols recommend radiation therapy (RT) both to the primary site and to sites of metastatic disease that persist after induction chemotherapy. Although there are abundant data showing excellent local control (LC) with 21 Gy directed at the primary site, there are few data describing the feasibility and efficacy of RT directed at metastatic sites of disease as part of consolidation. METHODS AND MATERIALS: All patients with neuroblastoma who received RT to metastatic sites of disease as a part of consolidative therapy at a single institution between 2000 and 2015 were reviewed. Among 159 patients, 244 metastases were irradiated. RESULTS: The median follow-up period among surviving patients was 7.4 years. Over 85% of the irradiated metastases were treated with 21 Gy (range, 10.5-36 Gy). Tumor recurrence occurred in 43 of 244 irradiated metastases (18%). The 5-year LC rate of treated metastatic sites was 81%. Metastatic sites that cleared with induction chemotherapy had improved LC compared with sites with persistent uptake on metaiodobenzylguanidine scans (LC rate, 92% vs 67%; P < .0001). LC at irradiated metastatic sites did not differ based on total number of sites irradiated or site of disease irradiated (bone vs soft tissue). Patients with bulky, resistant disease who were treated with 30 to 36 Gy had worse LC (P = .02). However, on multivariate analysis, only persistence after induction chemotherapy remained a significant prognostic factor for LC (hazard ratio, 3.7; P < .0001). Patients who had LC at irradiated metastatic sites had improved overall survival compared with those who did not (overall survival rate, 71% vs 50%; P < .0001). CONCLUSIONS: Response to chemotherapy is an important prognostic factor for LC at irradiated metastatic sites in neuroblastoma. Overall, consolidative RT appears to be an effective modality of LC. Long-term disease control can be achieved with such an approach.

Silk fibroin-Thelebolan matrix: A promising chemopreventive scaffold for soft tissue cancer.

Research of improved functional bio-mimetic matrix for regenerative medicine is currently one of the rapidly growing fields in tissue engineering and medical sciences. This study reports a novel bio-polymeric matrix, which is fabricated using silk protein fibroin from Bombyx mori silkworm and fungal exopolysaccharide Thelebolan from Antarctic fungus Thelebolus sp. IITKGP-BT12 by solvent evaporation and freeze drying method. Natural cross linker genipin is used to imprison the Thelebolan within the fibroin network. Different cross-linked and non-cross-linked fibroin/Thelebolan matrices are fabricated and biophysically characterized. Cross-linked thin films show robustness, good mechanical strength and high temperature stability in comparison to non-cross-linked and pure matrices. The 3D sponge matrices demonstrate good cytocompatibility. Interestingly, sustained release of the Thelebolan from the cross-linked matrices induce apoptosis in colon cancer cell line (HT-29) in time dependent manner while it is nontoxic to the normal fibroblast cells (L929).The findings indicate that the cross-linked fibroin/Thelebolan matrices can be used as potential topical chemopreventive scaffold for preclusion of soft tissue carcinoma.

The role of margins in extremity soft tissue sarcoma.

For extremity soft tissue sarcomas, limb salvage is now standard of care. The extent of surgical margins is balanced with functionality of the resected limb. Although negative margins are the goal, the necessary width is unclear. Additional considerations for margin adequacy include presence of anatomic barriers such as fascia and periosteum, proximity of critical structures, receipt of adjuvant and neoadjuvant therapies, and histologic subtype. Multidisciplinary team discussion is critical for treatment planning.

Fibrosis and hypoxia-inducible factor-1α-dependent tumors of the soft tissue on loss of von Hippel-Lindau in mesenchymal progenitors.

The hypoxia-inducible factor (Hif)-1α (Hif-1α) and Hif-2α (Epas1) have a critical role in both normal development and cancer. von Hippel Lindau (Vhl) protein, encoded by a tumor suppressor gene, is an E3 ubiquitin ligase that targets Hif-1α and Epas1 to the proteasome for degradation. To better understand the role of Vhl in the biology of mesenchymal cells, we analyzed mutant mice lacking Vhl in mesenchymal progenitors that give rise to the soft tissues that form and surround synovial joints. Loss of Vhl in mesenchymal progenitors of the limb bud caused severe fibrosis of the synovial joints and formation of aggressive masses with histologic features of mesenchymal tumors. Hif-1α and its downstream target connective tissue growth factor were necessary for the development of these tumors, which conversely still developed in the absence of Epas1, but at lower frequency. Human tumors of the soft tissue are a very complex and heterogeneous group of neoplasias. Our novel findings in genetically altered mice suggest that activation of the HIF signaling pathway could be an important pathogenetic event in the development and progression of at least a subset of these tumors.

Sarcomas and the immune system: implications for therapeutic strategies.

Soft-tissue sarcomas are a heterogeneous group of tumors that are capable of generating host immune responses. Historically the role of antitumor immunity was first studied in soft-tissue sarcomas. Subsequent in vitro studies, preclinical models, and clinical observations have provided ample evidence for an immunologic approach to sarcoma treatment. Initial clinical trials involving vaccines and adoptive immunotherapy have demonstrated promising results. The continued search for sarcoma tumor-associated antigens as specific targets is central to the clinical translation of effective immunotherapies.

Follow-up after primary treatment of soft tissue sarcoma of extremities: impact of frequency of follow-up imaging on disease-specific survival.

BACKGROUND AND OBJECTIVES: We explored the impact of frequency of surveillance imaging on disease-specific survival (DSS) in patients with extremity soft tissue sarcoma (STS). METHODS: Locoregional imaging (LRI) and chest imaging (CI) were used to detect local recurrence (LR) and distant metastasis (DM), respectively. Relapsing patients were retrospectively assigned to more frequent surveillance (MFS) or less frequent surveillance (LFS) groups, according to the median interval for each follow-up modality. Outcome measures included overall DSS (O-DSS), post-LR DSS, and post-DM DSS. RESULTS: We assigned 165 patients to three distinct risk groups according to tumor size (≤5 vs. >5 cm), depth (superficial- vs. deep-seated), grade (I vs. II or III), and surgical margin (≥10 vs. <10 mm). Data for 80 patients who relapsed were analyzed. Among 50 high-risk (with all four risk factors) relapsing patients, those in the MFS group for either LRI or CI had better O-DSS (LRI, median 44.07 vs. 27.43 months, P = 0.008; CI, median 43.60 vs. 36.93 months, P = 0.036), post-LR DSS (median 27.20 vs. 10.63 months, P = 0.028) and post-DM DSS (median 13.20 vs. 6.24 months, P = 0.031). CONCLUSION: More frequent follow-up were associated with improved survival in high-risk relapsing patients with extremity STS by providing greater opportunities for adequate reoperation.

Adjuvant chemotherapy decreases and postpones distant metastasis in extremity stage IIB/III synovial sarcoma patients.

BACKGROUND AND OBJECTIVES: The role of adjuvant chemotherapy (AC) in treatment of synovial sarcoma remains controversial. Aim of our study is to investigate the influence of AC on disease-specific survival and metastasis free survival, the difference in time to metastasis (TTM) was also analyzed. MATERIALS AND METHODS: Seventy-six cases of stage IIB/III synovial sarcoma from January 1993 to December 2008 were retrospectively analyzed. All the 76 patients were treated with surgical resection. AC regimen included first line MAID or AIM, second line Gemcitabine + Docetaxel with sufficient dose intensity. The clinical, pathologic, and treatment variables were analyzed for disease specific survival (DSS), metastasis free survival (MFS) and TTM. RESULTS: Median follow up period was 68 months. 51 patients received AC (51/76, 67%), 25 received no adjuvant chemotherapy (NAC, 25/76, 33%). The 5-year DSS of the AC patients was 73%(58-87%) compared with 31%(19-52%) for the NAC patients (P = 0.001). AC was also independently associated with improved MFS (P = 0.008) and prolonged TTM (P = 0.001). CONCLUSIONS: Patients with stage IIB/III synovial sarcoma might benefit in DSS, MFS, and a prolonged TTM from AC.

[Forefoot and midfoot amputations]. / Amputationen an Vor- und Mittelfuß

OBJECTIVE: Partial foot amputations are feasible regardless of the causal condition, including peripheral vascular disease with a few exceptions. Compared to higher amputation levels, a good foot stump permits full end bearing and enables the patient, even with a hindfoot stump, to walk without the need for a prosthesis. The more peripheral the amputation level selected, the greater the need for gentle tissue handling and meticulous postoperative care, but also the greater the risk of a breakdown requiring stump revision surgery. In the forefoot, partial amputation of the metatarsals preserves the length of the stump and, thus, minimizes the loss of weight-bearing surface. The resection of metatarsal and midfoot bones without removing the toes, called a "hidden" amputation, is more acceptable to the patient who does not feel as if he/she has become an amputee. In addition, no neuroma or phantom pain is experienced. Biomechanically, this amputation hardly differs from a classical amputation. INDICATIONS: Amputation cannot be avoided by any conservative or operative means. CONTRAINDICATIONS: Absolute: rapidly progressing peripheral arterial diseases, i.e., Buerger-Winiwarter's disease. Relative: renal failures requiring dialysis or kidney transplantation. SURGICAL TECHNIQUE: Patient in prone position, keep foot and calf free, protect heel from pressure. Mark the skin incisions. A long plantar flap covers the bones and is sutured to the short dorsal flap at the dorsum of the foot. Begin with the dorsal incision down to the bones. After separating the bones, turn the distal part down and separate the plantar soft tissue flap. The bones are well aligned and shaped. Longitudinal amputations preserve a larger load-bearing surface and, therefore, are preferred, if possible. Another alternative is the "hidden" amputation. Except for amputations in peripheral vascular diseases, the digits and their neurovascular supplies are preserved. Only the bones are resected, from transmetatarsal to Chopart. The toes will retract within a few weeks. The patients do not feel as if she/he has become an amputee, although the biomechanics of the foot are about the same as after a total amputation. In case of infection, wound debridement, open wound treatment, and delayed primary closure are recommended. POSTOPERATIVE MANAGEMENT: Full plantar weight bearing in plaster cast or walker is possible 4-6 weeks after surgery. In the case of diabetic foot, healing can require weeks. Total contact prosthesis without limiting the range of motion (ROM) of the ankle and the subtalar joint after 6 weeks. Best results are obtained with prostheses using the silicone technique. Alternative: orthopedic footwear. RESULTS: It is desirable to maintain the greatest length possible; wound healing disorders are observed in over half of all cases. Operative stump corrections are justified in 20-30%; a transtibial amputation is seldom necessary.

A direct role for Met endocytosis in tumorigenesis.

Compartmentalization of signals generated by receptor tyrosine kinase (RTK) endocytosis has emerged as a major determinant of various cell functions. Here, using tumour-associated Met-activating mutations, we demonstrate a direct link between endocytosis and tumorigenicity. Met mutants exhibit increased endocytosis/recycling activity and decreased levels of degradation, leading to accumulation on endosomes, activation of the GTPase Rac1, loss of actin stress fibres and increased levels of cell migration. Blocking endocytosis inhibited mutants' anchorage-independent growth, in vivo tumorigenesis and metastasis while maintaining their activation. One mutant resistant to inhibition by a Met-specific tyrosine kinase inhibitor was sensitive to endocytosis inhibition. Thus, oncogenicity of Met mutants results not only from activation but also from their altered endocytic trafficking, indicating that endosomal signalling may be a crucial mechanism regulating RTK-dependent tumorigenesis.

Metronomic therapy with cyclophosphamide and piroxicam effectively delays tumor recurrence in dogs with incompletely resected soft tissue sarcomas.

BACKGROUND: Continuous administration of low doses of cyclophosphamide and standard doses of cyclooxygenase-inhibiting drugs has been shown to suppress tumor angiogenesis, reverse immunosuppression, and deplete regulatory T cells in cancer models. HYPOTHESIS: We hypothesized that continuous treatment with low-dose cyclophosphamide and full-dose piroxicam would delay tumor recurrence in dogs with soft tissue sarcomas (STS). ANIMALS: Eighty-five dogs with incompletely resected STS, 30 treated dogs, and 55 contemporary control dogs. METHODS: Treatment outcomes in 85 dogs with incompletely resected STS were evaluated in a retrospective study. Dogs in the treatment group received continuously administered low-dose cyclophosphamide (10 mg/m2) and standard dose piroxicam (0.3 mg/kg) therapy. Time to local tumor recurrence (disease-free interval; DFI) was compared between the 30 treated dogs and 55 untreated control dogs matched for age and tumor site and grade. RESULTS: DFI was significantly (P < .0001) prolonged for STS of all sites (trunk and extremity) in treated dogs compared with untreated control dogs. The DFI also was significantly longer in treated dogs when tumor site (trunk and extremity) was compared. Twelve treated dogs (40%) experienced mild toxicity (grade 1 and 2) at some point during treatment and 1 dog developed grade 4 cystitis. Every other day dosing was tolerated better than daily dosing. CONCLUSIONS: Metronomic therapy with cyclophosphamide and piroxicam was very effective in preventing tumor recurrence in dogs with incompletely resected STS. These findings suggest that further evaluation of this approach is warranted as adjuvant therapy in dogs with highly metastatic tumors such as osteosarcoma and melanoma.

Prognostic Factors for Injection-Site Sarcomas in Cats.

In this retrospective study of cases from Italy and Switzerland, the clinical features of injection-site sarcomas in 57 cats were reviewed. In addition to routine physical, laboratory, and radiographic examinations, advanced imaging (computed tomographic scans) was conducted to delineate tumor extent. Treatment was by wide surgical excision including partial scapulectomy, spinous process removal, and amputation (appendicular lesions). Radiotherapy and chemotherapy were also used. The results revealed that cats with grade 3 tumors were most likely to develop distant pulmonary metastases (and to have shorter survival rates); this occurred in 21% of the cases.Median follow-up for cats that survived (51%) was 600 days; for those that died (49%), it was 272 days. In this study, tumor size, as measured manually by calipers, was not a prognostic factor. Furthermore, factors associated with local recurrence were not identified. The authors recognized the limitations of a retrospective study, especially with regard to treatments before referral.

A non-RGD-based integrin binding peptide (ATN-161) blocks breast cancer growth and metastasis in vivo.

PURPOSE: Integrins are expressed by numerous tumor types including breast cancer, in which they play a crucial role in tumor growth and metastasis. In this study, we evaluated the ability of ATN-161 (Ac-PHSCN-NH2), a 5-mer capped peptide derived from the synergy region of fibronectin that binds to alpha5beta1 and alphavbeta3 in vitro, to block breast cancer growth and metastasis. EXPERIMENTAL DESIGN: MDA-MB-231 human breast cancer cells were inoculated s.c. in the right flank, or cells transfected with green fluorescent protein (MDA-MB-231-GFP) were inoculated into the left ventricle of female BALB/c nu/nu mice, resulting in the development of skeletal metastasis. Animals were treated with vehicle alone or by i.v. infusion with ATN-161 (0.05-1 mg/kg thrice a week) for 10 weeks. Tumor volume was determined at weekly intervals and tumor metastasis was evaluated by X-ray, microcomputed tomography, and histology. Tumors were harvested for histologic evaluation. RESULT: Treatment with ATN-161 caused a significant dose-dependent decrease in tumor volume and either completely blocked or caused a marked decrease in the incidence and number of skeletal as well as soft tissue metastases. This was confirmed histologically as well as radiographically using X-ray and microcomputed tomography. Treatment with ATN-161 resulted in a significant decrease in the expression of phosphorylated mitogen-activated protein kinase, microvessel density, and cell proliferation in tumors grown in vivo. CONCLUSION: These studies show that ATN-161 can block breast cancer growth and metastasis, and provides a rationale for the clinical development of ATN-161 for the treatment of breast cancer.

High-dose radioimmunotherapy combined with fixed, low-dose paclitaxel in metastatic prostate and breast cancer by using a MUC-1 monoclonal antibody, m170, linked to indium-111/yttrium-90 via a cathepsin cleavable linker with cyclosporine to prevent human anti-mouse antibody.

PURPOSE: Although radioimmunotherapy alone is effective in lymphoma, its application to solid tumors will likely require a combined modality approach. In these phase I studies, paclitaxel was combined with radioimmunotherapy in patients with metastatic hormone-refractory prostate cancer or advanced breast cancer. EXPERIMENTAL DESIGN: Patients were imaged with indium-111 (111In)-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid-peptide-m170. One week later, yttrium-90 (90Y)-m170 was infused (12 mCi/m2 for prostate cancer and 22 mCi/m2 for breast cancer). Initial cohorts received radioimmunotherapy alone. Subsequent cohorts received radioimmunotherapy followed 48 hours later by paclitaxel (75 mg/m2). Cyclosporine was given to prevent development of human anti-mouse antibody. RESULTS: Bone and soft tissue metastases were targeted by 111In-m170 in 15 of the 16 patients imaged. Three prostate cancer patients treated with radioimmunotherapy alone had no grade 3 or 4 toxicity. With radioimmunotherapy and paclitaxel, two of three prostate cancer patients developed transient grade 4 neutropenia. Four breast cancer patients treated with radioimmunotherapy alone had grade 3 or 4 myelosuppression. With radioimmunotherapy and paclitaxel, both breast cancer patients developed grade 4 neutropenia. Three breast cancer patients required infusion of previously harvested peripheral blood stem cells because of neutropenic fever or bleeding. One patient in this trial developed human anti-mouse antibody in contrast to 12 of 17 patients in a prior trial using m170-radioimmunotherapy without cyclosporine. CONCLUSIONS: 111In/90Y-m170 targets prostate and breast cancer and can be combined with paclitaxel with toxicity limited to marrow suppression at the dose levels above. The maximum tolerated dose of radioimmunotherapy and fixed-dose paclitaxel with peripheral blood stem cell support has not been reached. Cyclosporine is effective in preventing human anti-mouse antibody, suggesting the feasibility of multidose, "fractionated" therapy that could enhance clinical response.

Significant reduction of laparotomy-associated lung metastases and subcutaneous tumors after perioperative immunomodulation with flt3 ligand in mice.

Laparotomy has been associated with increased rates of tumor establishment and metastasis formation postoperatively in animal models. The purpose of this study was to determine the impact on postoperative tumor growth of perioperative upregulation of immune function via fetal liver tyrosine kinase 3 (Flt3 ligand). Two murine studies were carried out: the first utilized a lung metastases model, and the second involved a subcutaneous tumor model. Each study included four groups: anesthesia control (AC), AC plus Flt3 ligand (ACFlt3), sham laparotomy (OP), and OP plus Flt3 ligand (OPFlt3). Flt3 ligand was administered by daily intraperitoneal injection (10 mug/dose) beginning 5 days preoperatively and continuing for 1 week postoperatively. In study 1, A/J mice were given tail vein injections of 1.5 x 10(5) TA3Ha cancer cells on the day of surgery. The mice were sacrificed 14 days after surgery, the lungs processed, and the surface metastases counted by a blinded observer. In study 2 C3H/He mice were given a dorsal subcutaneous injection of 10(4) MC-2 cancer cells on the day of surgery. The mice were sacrificed 31 days after surgery, and the injection sites were evaluated for subcutaneous tumors grossly and histologically. In study 1, the median number of surface lung metastases per mouse was 166 in the OP group and 38 in the OPFlt3 (P = .021). Mice in the AC group developed a median 50 lung metastases per animal compared with mice in the ACFlt3 group who had a median of 10 metastases per mouse (P = .001). The OP group had significantly more metastases than the AC group (P = .048). In study 2, the percentage of animals that developed tumors in the AC, OP, ACFlt3, and OPFlt3 groups was 43, 80, 0, and 20, respectively. The incidence of tumors in the OPFLt3 group and the ACFlt3 group was significantly less than their respective control groups (P < .01). The difference between the OP and AC groups was not significant (P > .05). Perioperatively administered Flt3 ligand was associated with significantly fewer lung metastases and a lower incidence of subcutaneous tumor formation after laparotomy and anesthesia alone. Perioperative immunomodulation may limit untoward surgery-related oncologic effects.

Ineffectiveness of dietary folic acid supplementation on the incidence of lipomyelomeningocele: pathogenetic implications.

OBJECT: Periconceptual folic acid supplementation is effective in myelomeningocele prevention. The relationship between folic acid and lipomyelomeningocele (LMM) and the overall incidence of this occult form of spina bifida has never been studied. The objectives of this study were to determine the impact of dietary folic acid supplementation on the incidence of LMM and to measure its overall incidence. METHODS: In a retrospective population-based study the authors calculated the incidence of LMM in Nova Scotia between 1985 and 2001. Because of changes in public policy during this period, there are three intervals defined in relation to the treatment of the food supply with folic acid: 1) prior to folic acid fortification (1985-1994); 2) postsupplementation but prefortification (1995-1998); and 3) postfortification. The overall incidence of LMM in Nova Scotia between 1985 and 2001 was 16 per 100,000 live births or one case per 6121 live births. Its incidence between 1985 and 1994 was 15 per 100,000 live births, and between 1995 and 1998 it was 12 per 100.000 live births (relative risk [RR] 0.82, 95% confidence interval [CI] 0.31-2.22; p = 0.7). Between 1999 and 2001, the incidence of LMM was 29 per 100,000 live births, which was not significantly different from that between 1995 and 1998 (RR 2.41. 95% CI 0.79-7.36; p = 0.11) or between 1985 and 1994 (RR 1.98, 95% CI 0.86-4.56; p = 0.1). CONCLUSIONS: The overall incidence of LMM between 1985 and 2001 in Nova Scotia was 16 per 100,000 live births and has not been reduced by dietary folic acid supplementation. This finding provides epidemiological evidence that the embryogenesis of LMM is fundamentally different from that of myelomeningocele.