Biological and therapeutic insights from animal modeling of fusion-driven pediatric soft tissue sarcomas.

Survival for children with cancer has primarily improved over the past decades due to refinements in surgery, radiation and chemotherapy. Although these general therapies are sometimes curative, the cancer often recurs, resulting in poor outcomes for patients. Fusion-driven pediatric soft tissue sarcomas are genetically defined by chromosomal translocations that create a chimeric oncogene. This distinctive, almost 'monogenic', genetic feature supports the generation of animal models to study the respective diseases in vivo. This Review focuses on a subset of fusion-driven pediatric soft tissue sarcomas that have transgenic animal tumor models, which includes fusion-positive and infantile rhabdomyosarcoma, synovial sarcoma, undifferentiated small round cell sarcoma, alveolar soft part sarcoma and clear cell sarcoma. Studies using the animal models of these sarcomas have highlighted that pediatric cancers require a specific cellular state or developmental stage to drive tumorigenesis, as the fusion oncogenes cause different outcomes depending on their lineage and timing of expression. Therefore, understanding these context-specific activities could identify targetable activities and mechanisms critical for tumorigenesis. Broadly, these cancers show dependencies on chromatin regulators to support oncogenic gene expression and co-opting of developmental pathways. Comparative analyses across lineages and tumor models will further provide biological and therapeutic insights to improve outcomes for these children.

Effect of high-energy mechanical milling on the physicochemical and rheological properties of chayotextle (Sechium edule Sw.) starch.

This work evaluates the effect of high-energy mechanical milling time (7 levels, 20-80 min) on amylose content, crystallinity pattern, temperature and gelatinization enthalpy, morphology, and rheological properties of chayotextle (Sechium edule Sw.) starch. After 30 min of milling, granular structure was affected, and amylose values were the highest while crystallinity and gelatinization enthalpy decreased significantly. These changes allowed to obtain gels with viscoelastic properties where the elastic character (Ç´) prevailed upon the viscous modulus (Ǵ́). Native starch showed Tan δ values of 0.6, increased significantly (0.9) after 30 min of milling due to the surge in linear chains (amylose) and loss of granular structure. Native and modified starches showed high dependence on cutting or shear speed, presenting a non-Newtonian behavior (reofluidizers). These results indicate that mechanical grinding is an alternative to obtain modified starches with applications in the food industry.

Functionality and characterization of modified starch films with pineapple leaf fibers.

The objective of this work was to modify banana starch with pineapple leaf fibers (PALF) and its production of biodegradable films. The reaction conditions of the starch modification were a Starch/PALF mass ratio of 50, a time of 1 h and a temperature of 140 °C, to obtain a yield of 41.18 %. Characterization by FTIR and NMR confirmed that the chemical reaction was carried out. XRD and TGA analysis showed that the crystalline zones of the starch were affected during the modification and the product obtained is thermally less stable compared to unmodified starch. The modified starch showed a lower pasting profile compared to the native starch; however, the modified starch showed the ability to form a film. The starch-PALF films were obtained by the casting method and partially characterized. These films presented better mechanical properties compared to the unmodified films. Also, these films could compete with conventional non-biodegradable plastics.

VGLL2-NCOA2 leverages developmental programs for pediatric sarcomagenesis.

Clinical sequencing efforts are rapidly identifying sarcoma gene fusions that lack functional validation. An example is the fusion of transcriptional coactivators, VGLL2-NCOA2, found in infantile rhabdomyosarcoma. To delineate VGLL2-NCOA2 tumorigenic mechanisms and identify therapeutic vulnerabilities, we implement a cross-species comparative oncology approach with zebrafish, mouse allograft, and patient samples. We find that VGLL2-NCOA2 is sufficient to generate mesenchymal tumors that display features of immature skeletal muscle and recapitulate the human disease. A subset of VGLL2-NCOA2 zebrafish tumors transcriptionally cluster with embryonic somitogenesis and identify VGLL2-NCOA2 developmental programs, including a RAS family GTPase, ARF6. In VGLL2-NCOA2 zebrafish, mouse, and patient tumors, ARF6 is highly expressed. ARF6 knockout suppresses VGLL2-NCOA2 oncogenic activity in cell culture, and, more broadly, ARF6 is overexpressed in adult and pediatric sarcomas. Our data indicate that VGLL2-NCOA2 is an oncogene that leverages developmental programs for tumorigenesis and that reactivation or persistence of ARF6 could represent a therapeutic opportunity.

Zebrafish her3 knockout impacts developmental and cancer-related gene signatures.

HES3 is a basic helix-loop-helix transcription factor that regulates neural stem cell renewal during development. HES3 overexpression is predictive of reduced overall survival in patients with fusion-positive rhabdomyosarcoma, a pediatric cancer that resembles immature and undifferentiated skeletal muscle. However, the mechanisms of HES3 cooperation in fusion-positive rhabdomyosarcoma are unclear and are likely related to her3/HES3's role in neurogenesis. To investigate HES3's function during development, we generated a zebrafish CRISPR/Cas9 null mutation of her3, the zebrafish ortholog of HES3. Loss of her3 is not embryonic lethal and adults exhibit expected Mendelian ratios. Embryonic her3 zebrafish mutants exhibit dysregulated neurog1 expression, a her3 target gene, and the mutant her3 fails to bind the neurog1 promoter sequence. Further, her3 mutants are significantly smaller than wildtype and a subset present with lens defects as adults. Transcriptomic analysis of her3 mutant embryos indicates that genes involved in organ development, such as pctp and grinab, are significantly downregulated. Further, differentially expressed genes in her3 null mutant embryos are enriched for Hox and Sox10 motifs. Several cancer-related gene pathways are impacted, including the inhibition of matrix metalloproteinases. Altogether, this new model is a powerful system to study her3/HES3-mediated neural development and its misappropriation in cancer contexts.

Modification of banana starch (Musa paradisiaca L.) with polyethylene terephthalate: Virgin and bottle waste.

Chemical modification of banana starch (Musa paradisiaca L.) with the degradation products of virgin and bottle waste of polyethylene terephthalate was carried out in situ. The modified starch was characterized by FTIR and NMR, which allowed proposing three chemical structures. SEM micrographs showed that the morphology of the modified starch granule is directly related with mass ratio of Starch/PET and type of PET used in the reaction. The crystallinity of the modified starch decreased up to 92.6% and 62.5% using bottle waste and virgin PET, respectively, according to XRD diffractograms. TGA analysis showed that the starch degradation temperature decreased by 12 °C. Modified starch films were elaborate and its electrical conductivity was found to be 2.9 times compared to that of native starch. The starch/PET film presented the highest value in the mechanical property of elongation at break compared to the starch-only film. The modified starch film was degraded above 80% by aqueous hydrolysis.

Sporadic activation of an oxidative stress-dependent NRF2-p53 signaling network in breast epithelial spheroids and premalignancies.

Breast and mammary epithelial cells experience different local environments during tissue development and tumorigenesis. Microenvironmental heterogeneity gives rise to distinct cell regulatory states whose identity and importance are just beginning to be appreciated. Cellular states diversify when clonal three-dimensional (3D) spheroids are cultured in basement membrane, and one such state is associated with stress tolerance and poor response to anticancer therapeutics. Here, we found that this state was jointly coordinated by the NRF2 and p53 pathways, which were costabilized by spontaneous oxidative stress within 3D cultures. Inhibition of NRF2 or p53 individually disrupted some of the transcripts defining the regulatory state but did not yield a notable phenotype in nontransformed breast epithelial cells. In contrast, combined perturbation prevented 3D growth in an oxidative stress-dependent manner. By integrating systems models of NRF2 and p53 signaling in a single oxidative stress network, we recapitulated these observations and made predictions about oxidative stress profiles during 3D growth. NRF2 and p53 signaling were similarly coordinated in normal breast epithelial tissue and hormone-negative ductal carcinoma in situ lesions but were uncoupled in triple-negative breast cancer (TNBC), a subtype in which p53 is usually mutated. Using the integrated model, we correlated the extent of this uncoupling in TNBC cell lines with the importance of NRF2 in the 3D growth of these cell lines and their predicted handling of oxidative stress. Our results point to an oxidative stress tolerance network that is important for single cells during glandular development and the early stages of breast cancer.

Thermal, morphological and structural characterization of a copolymer of starch and polyethylene.

The objective of this paper was to perform a copolymerization between polyethylene and starch in order to obtain new environmentally friendly materials. The copolymer obtained was characterized thermally, morphologically and structurally, including its pasting profile. The starch-g-PE copolymer showed lower thermal stability compared to the control materials. FTIR analysis determined that the chemical bond signal between the starch and polyethylene in the copolymer overlaps with the native starch signals. The signal from this chemical bond was assigned by proton NMR spectroscopy at δ 4.45 ppm. X-ray studies of the copolymer showed a material with more amorphous characteristics compared to native starch. SEM analysis demonstrated the presence of cracks in the starch granules which favored the chemical interaction between the polymers. The pasting behavior of the copolymer was less pronounced compared to native starch. Therefore, the copolymerization of both polymers could be an alternative to recycle polyethylene and make biodegradable materials.