Springer protocols feed by Cancer Research 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Detecting PTEN and PI3K Signaling in Brain
The central nervous system is comprised of multiple cell types including neurons, glia, and other supporting cells that may differ dramatically in levels of signaling pathway activation. Immunohistochemistry in conjunction with drug interference are powerful tools that allow evaluation of signaling pathways in different cell types of the mouse central nervous system in vivo. Here we provide detailed protocols for immunohistochemistry to evaluate three essential components in the PI3K pathway in mouse brain: Pten, p-Akt, and p-4ebp1, and for rapamycin treatment to modulate mTOR signaling in vivo. (Source: Springer protocols...
Source: Springer protocols feed by Cancer Research - November 18, 2015 Category: Cancer & Oncology Source Type: news
Measurement of PTEN by Flow Cytometry
Recent advancements have driven the development of smaller footprint, less expensive, and user-friendly flow cytometers introducing the technology to more users. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 18, 2015 Category: Cancer & Oncology Source Type: news
Immunopathologic Assessment of PTEN Expression
Immunohistochemistry (IHC) is an excellent technique used routinely to define the phenotype in pathology laboratories through the analysis of molecular expression in cells and tissues. The PTEN protein is ubiquitously expressed in the majority of human tissues, and allelic or complete loss of PTEN is frequently observed in different types of malignancies leading to an activation of the AKT/mTOR pathways. IHC-based analyses are best to determine the level of PTEN expression in histological samples, but not to assess partial or heterozygous deletions, for which FISH analyses are more appropriate. Interpretation of the IHC re...
Source: Springer protocols feed by Cancer Research - November 18, 2015 Category: Cancer & Oncology Source Type: news
PTEN at 18: Still Growing
Discovered in 1997, PTEN remains one of the most studied tumor suppressors. In this issue of Methods in Molecular Biology, we assembled a series of papers describing various clinical and experimental approaches to studying PTEN function. Due to its broad expression, regulated subcellular localization, and intriguing phosphatase activity, methodologies aimed at PTEN study have often been developed in the context of mutations affecting various aspects of its regulation, found in patients burdened with PTEN loss-driven tumors. PTEN’s extensive posttranslational modifications and dynamic localization pose unique challeng...
Source: Springer protocols feed by Cancer Research - November 18, 2015 Category: Cancer & Oncology Source Type: news
PTEN: History of a Tumor Suppressor
Starting from the discovery of “inhibitory chromosomes” by Theodor Boveri to the finding by Henry Harris that fusing a normal cell to a cancer cell reduced tumorigenic potential, the notion of tumor suppression was recognized well before any tumor-suppressor genes were discovered. Although not the first to be revealed, PTEN has been demonstrated to be one of the most frequently altered tumor suppressors in cancer. This introductory chapter provides a historical perspective on our current understanding of PTEN including some of the seminal discoveries in the tumor suppressor field, the events leading to PTEN&rsq...
Source: Springer protocols feed by Cancer Research - November 18, 2015 Category: Cancer & Oncology Source Type: news
Methods to Study PTEN in Mitochondria and Endoplasmic Reticulum
Although PTEN has been widely described as a nuclear and cytosolic protein, in the last 2 years, alternative organelles, such as the endoplasmic reticulum (ER), pure mitochondria, and mitochondria-associated membranes (MAMs), have been recognized as pivotal targets of PTEN activity. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 17, 2015 Category: Cancer & Oncology Source Type: news
Methods for PTEN in Stem Cells and Cancer Stem Cells
PTEN (phosphatase and tensin homologue) is the first tumor suppressor identified to have phosphatase activity and its gene is the second most frequently deleted or mutated tumor-suppressor gene associated with human cancers. Germline PTEN mutations are the cause of three inherited autosomal dominant disorders. Phosphatidylinositol 3,4,5,-triphosphate (PIP3), the product of the PI3 kinase, is one of the key intracellular targets of PTEN’s phosphatase activity, although PTEN’s phosphatase-independent activities have also been identified. PTEN is critical for stem cell maintenance, which contributes to its control...
Source: Springer protocols feed by Cancer Research - November 17, 2015 Category: Cancer & Oncology Source Type: news
Methods in the Study of PTEN Structure: X-Ray Crystallography and Hydrogen Deuterium Exchange Mass Spectrometry
Despite its small size and deceptively simple domain organization, PTEN remains a challenging structural target due to its N- and C-terminal intrinsically disordered segments, and the conformational heterogeneity caused by phosphorylation of its C terminus. Using hydrogen/deuterium exchange mass spectrometry (HDX-MS), it is possible to probe the conformational dynamics of the disordered termini, and also to determine how PTEN binds to lipid membranes. Here, we describe how to purify recombinant, homogenously dephosphorylated PTEN from a eukaryotic system for subsequent investigation with HDX-MS or crystallography. (Source:...
Source: Springer protocols feed by Cancer Research - November 17, 2015 Category: Cancer & Oncology Source Type: news
Modeling Cancer-Associated Mutations of PTEN in Mice
Manipulation of mammalian genome and generation of genetically engineered mouse models (GEMMs) has revolutionized the scientific approach to address biological questions. To date, a number of gene-targeting strategies have been devised and are available to investigators for the generation of genetically modified mouse lines. Nevertheless, irrespective of the methodological approach selected, there remain critical molecular steps that need to be performed and put in place in order to obtain controlled and well-characterized new animal models. Here we provide technical details for the (1) handling and maintenance of mouse em...
Source: Springer protocols feed by Cancer Research - November 17, 2015 Category: Cancer & Oncology Source Type: news
C. elegans Methods to Study PTEN
C. elegans encodes a PTEN homolog called DAF-18 and human PTEN can functionally replace DAF-18. Thus C. elegans provides a valuable model organism to study PTEN. This chapter provides methods to study DAF-18/PTEN function in C. elegans. We provide methods to genotype daf-18/Pten mutants, visualize and quantify DAF-18/PTEN in C. elegans, as well as to study physiological and developmental processes that will provide molecular insight on DAF-18/PTEN function. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 17, 2015 Category: Cancer & Oncology Source Type: news
Methods for the Identification of PTEN-Targeting MicroRNAs
The identification of PTEN-targeting microRNAs usually starts from an in silico bioinformatic prediction and then requires a careful experimental validation that exploits both heterologous and endogenous systems. Here we describe the methods used to carry on these analyses and experiments, examining pitfalls and alternatives for each step. Moreover, we give an overview of the latest high-throughput microRNA target identification techniques which offer a more comprehensive view of the microRNAs that can bind a fundamental tumor suppressor such as PTEN. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - November 17, 2015 Category: Cancer & Oncology Source Type: news
Clinical Evaluation of ErbB-Targeted CAR T-Cells, Following Intracavity Delivery in Patients with ErbB-Expressing Solid Tumors
Adoptive cell therapy using gene-modified T-cells has achieved impressive results in the treatment of B-cell malignancies. However, the development of similar strategies to treat solid tumors raises challenges with respect to tumor antigen selection and the achievement of efficient T-cell homing, survival and sustained effector function within the tumor microenvironment. To address these challenges, we have developed a gene-modified cellular therapy called T4 immunotherapy. To generate T4 immunotherapy, autologous T-cells are engineered by retroviral transduction to co-express two transgenes: (1) a chimeric antigen recepto...
Source: Springer protocols feed by Cancer Research - June 26, 2015 Category: Cancer & Oncology Source Type: news
Antiangiogenic Metargidin Peptide (AMEP) Gene Therapy in Disseminated Melanoma
Gene delivery by electroporation is an efficient method for transfecting genes into various tissues including tumors. Here we present the treatment protocol used in a phase 1 study on gene electrotransfer of plasmid DNA encoding an antiangiogenic peptide into cutaneous melanoma. (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - June 26, 2015 Category: Cancer & Oncology Source Type: news
A Phase 2, Open-Label, Randomized Study of Pexa-Vec (JX-594) Administered by Intratumoral Injection in Patients with Unresectable Primary Hepatocellular Carcinoma
We present the design of a randomized dose-finding trial of Pexa-Vec in patients with advanced HCC in which Pexa-Vec was delivered by intratumoral injection three times every 2 weeks at one of two dose levels (1 × 108 plaque forming units (pfu) versus 1 × 109 pfu). (Source: Springer protocols feed by Cancer Research)
Source: Springer protocols feed by Cancer Research - June 26, 2015 Category: Cancer & Oncology Source Type: news
Minicircle: Next Generation DNA Vectors for Vaccination
The use of DNA vaccines requires pharmaceutical grade DNA that causes the immunization on the basis of a nucleic acid sequence that encodes the protein to be vaccinated against. This nucleic acid sequence can be a circular or linear plasmid, preferably a double stranded one and should not contain any other and especially not any “toxic” sequences. Sequences that are not desirable to be part of the DNA drug can be those deriving from the (typically) bacterial amplification system to produce the DNA vaccine. These could be those portions of a plasmid that are only used for controlling the bacterial replication of...
Source: Springer protocols feed by Cancer Research - June 26, 2015 Category: Cancer & Oncology Source Type: news
