A Genetic Porcine Model of Cancer

The large size of the pig and its similarity in anatomy, physiology, metabolism, and genetics to humans make it an ideal platform to develop a genetically defined, large animal model of cancer. To this end, we created a transgenic oncopig line encoding Cre recombinase inducible porcine transgenes encoding KRASG12D and TP53R167H, which represent a commonly mutated oncogene and tumor suppressor in human cancers, respectively. Treatment of cells derived from these oncopigs with the adenovirus encoding Cre (AdCre) led to KRASG12D and TP53R167H expression, which rendered the cells transformed in culture and tumorigenic when engrafted into immunocompromised mice. Finally, injection of AdCre directly into these oncopigs led to the rapid and reproducible tumor development of mesenchymal origin. Transgenic animals receiving AdGFP (green fluorescent protein) did not have any tumor mass formation or altered histopathology. This oncopig line could thus serve as a genetically malleable model for potentially a wide spectrum of cancers, while controlling for temporal or spatial genesis, which should prove invaluable to studies previously hampered by the lack of a large animal model of cancer

The Potential of Nanotechnology in Medically Assisted Reproduction

Reproductive medicine is a field of science which searches for new alternatives not only to help couples achieve pregnancy and preserve fertility, but also to diagnose and treat diseases which can impair the normal operation of the reproductive tract. Assisted reproductive technology (ART) is a set of methodologies applied to cases related to infertility. Despite being highly practiced worldwide, ART presents some challenges, which still require special attention. Nanotechnology, as a tool for reproductive medicine, has been considered to help overcome some of those impairments. Over recent years, nanotechnology approaches applied to reproductive medicine have provided strategies to improve diagnosis and increase specificity and sensitivity. For in vitro embryo production, studies in non-human models have been used to deliver molecules to gametes and embryos. The exploration of nanotechnology for ART would bring great advances. In this way, experiments in non-human models to test the development and safety of new protocols using nanomaterials are very important for informing potential future employment in humans. This paper presents recent developments in nanotechnology regarding impairments still faced by ART: ovary stimulation, multiple pregnancy, and genetic disorders. New perspectives for further use of nanotechnology in reproductive medicine studies are also discussed

Análogo de curcumina induz superexpressão de caspases em linhagem de carcinoma de bexiga

O câncer se tornou uma epidemia em escala mundial. Dentre os vários tipos de neoplasia, o câncer de bexiga é o décimo mais frequente. Este tipo de tumor merece destaque pelo fato de representar o maior custo econômico em relação ao seu tratamento para a saúde pública, pelo fato de ter um longo período de tratamento. Visto isso, embora existam abordagens quimioterápicas comumente utilizadas na clínica para tratar neoplasias de bexiga, elas ainda apresentam inúmeros efeitos adversos. Assim, no âmbito do desenvolvimento de fármacos antitumorais, compostos sintéticos representam uma das principais opções no combate ao câncer. A curcumina é um composto extraído do açafrão-da-terra, com capacidade de atuar em várias doenças inclusive o câncer porém, sua utilização é limitada devido à baixa solubilidade e biodisponibilidade. Nesse contexto, devido ao potencial que os compostos sintéticos apresentam e à importância da contínua busca de moléculas com ação antitumoral, o presente trabalho teve como objetivo avaliar o potencial de indução de expressão de caspases pelos compostos análogos de curcumina 1a e 1d, além de analisar sua interação com receptores e proteínas relacionados com fatores de crescimento por meio de docagem molecular

Emerging technologies to create inducible and genetically defined porcine cancer models

There is an emerging need for new animal models that address unmet translational cancer research requirements. Transgenic porcine models provide an exceptional opportunity due to their genetic, anatomic and physiological similarities with humans. Due to recent advances in the sequencing of domestic animal genomes and the development of new organism cloning technologies, it is now very feasible to utilize pigs as a malleable species, with similar anatomic and physiological features with humans, in which to develop cancer models. In this review, we discuss genetic modification technologies successfully used to produce porcine biomedical models, in particular the Cre-loxP System as well as major advances and perspectives the CRISPR/Cas9 System. Recent advancements in porcine tumor modeling and genome editing will bring porcine models to the forefront of translational cancer research

The Oncopig Cancer Model as a Complementary Tool for Phenotypic Drug Discovery

The screening of potential therapeutic compounds using phenotypic drug discovery (PDD) is being embraced once again by researchers and pharmaceutical companies as an approach to enhance the development of new effective therapeutics. Before the genomics and molecular biology era and the consecutive emergence of targeted-drug discovery approaches, PDD was the most common platform used for drug discovery. PDD, also known as phenotypic screening, consists of screening potential compounds in either in vitro cellular or in vivo animal models to identify compounds resulting in a desirable phenotypic change. Using this approach, the biological targets of the compounds are not taken into consideration. Suitable animal models are crucial for the continued validation and discovery of new drugs, as compounds displaying promising results in phenotypic in vitro cell-based and in vivo small animal model screenings often fail in clinical trials. Indeed, this is mainly a result of differential anatomy, physiology, metabolism, immunology, and genetics between humans and currently used pre-clinical small animal models. In contrast, pigs are more predictive of therapeutic treatment outcomes in humans than rodents. In addition, pigs provide an ideal platform to study cancer due to their similarities with humans at the anatomical, physiological, metabolic, and genetic levels. Here we provide a mini-review on the reemergence of PDD in drug development, highlighting the potential of porcine cancer models for improving pre-clinical drug discovery and testing. We also present precision medicine based genetically defined swine cancer models developed to date and their potential as biomedical models

Summary of porcine tumor growth in vivo.

<p>^a Offspring sired by clone 63–3</p><p>^bIM, intramuscular; SQ, subcutaneous; IT, intra-testicular</p><p>^cConcentration of 2x10⁹ pfu AdCre/ml</p><p>^dSize 10 days post-injection (cm)</p><p>^eSize 20 days post-injection (cm)</p><p>^fRatio of normalized RNAseq levels between AdCre induced tumor tissue RNA/untreated transgenic muscle tissue RNA</p><p>Summary of porcine tumor growth in vivo.</p

Inducible expression of <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i> is transforming and tumorigenic.

<p>(<i>a</i>) RT-PCR analysis of <i>KRAS</i>^<i>G12D</i> and <i>TP53</i>^<i>R167H</i> mRNA expression in the fibroblast cell lines from each of the 4 transgenic clones treated with AdCre or AdGFP. (<i>b</i>) Comparison of cell morphology between AdCre and untreated control cells in culture, stained with H&E. (<i>c</i>) Normalized MFU measured by FACS at time points following Carboxyfluorescein succinimidyl ester (CFSE) dye loading of cells. (<i>d</i>) Graphical analysis of the mean number of migrating cells from triplicate plating of each of the 4 cell lines. (<i>e</i>) Graphical analysis of the mean number of colonies growing in soft agar for each cell line from triplicate plating. (<i>c-e</i>: all data points are the mean of the 4 cell lines derived from pigs 63–1, 63–2, 63–3, 63–4; error bars = SD; *p-value ≤ 0.05; **p-value ≤ 0.01).</p

Tumors arose from each AdCre treated cell line injected into immune compromised mice.

<p>Similar tumors (a) developed from all AdCre cell lines and no tumors developed from AdGFP cell injections; Histological analysis (b) revealed the tumors to be densely cellular non encapsulated and infiltrative neoplasm. 10x and insert 40X H&E Stain.</p

Injection of AdGFP into transgenic pigs did not induce tumors.

<p>The lack of any detectable changes to the injection site on d20 post- injection is shown at the surface (a–c); in underlying tissue (d–f); or upon histological examination (g–i).</p