Potential geothermal resources of Texas

UT Librarie

Gauging NOTCH1 Activation in Cancer Using Immunohistochemistry

Fixed, paraffin-embedded (FPE) tissues are a potentially rich resource for studying the role of NOTCH1 in cancer and other pathologies, but tests that reliably detect activated NOTCH1 (NICD1) in FPE samples have been lacking. Here, we bridge this gap by developing an immunohistochemical (IHC) stain that detects a neoepitope created by the proteolytic cleavage event that activates NOTCH1. Following validation using xenografted cancers and normal tissues with known patterns of NOTCH1 activation, we applied this test to tumors linked to dysregulated Notch signaling by mutational studies. As expected, frequent NICD1 staining was observed in T lymphoblastic leukemia/lymphoma, a tumor in which activating NOTCH1 mutations are common. However, when IHC was used to gauge NOTCH1 activation in other human cancers, several unexpected findings emerged. Among B cell tumors, NICD1 staining was much more frequent in chronic lymphocytic leukemia than would be predicted based on the frequency of NOTCH1 mutations, while mantle cell lymphoma and diffuse large B cell lymphoma showed no evidence of NOTCH1 activation. NICD1 was also detected in 38% of peripheral T cell lymphomas. Of interest, NICD1 staining in chronic lymphocytic leukemia cells and in angioimmunoblastic lymphoma was consistently more pronounced in lymph nodes than in surrounding soft tissues, implicating factors in the nodal microenvironment in NOTCH1 activation in these diseases. Among carcinomas, diffuse strong NICD1 staining was observed in 3.8% of cases of triple negative breast cancer (3 of 78 tumors), but was absent from 151 non-small cell lung carcinomas and 147 ovarian carcinomas. Frequent staining of normal endothelium was also observed; in line with this observation, strong NICD1 staining was also seen in 77% of angiosarcomas. These findings complement insights from genomic sequencing studies and suggest that IHC staining is a valuable experimental tool that may be useful in selection of patients for clinical trials

Session 13: Technology Transfer of Geopressured/Geothermal Resources to Industry

This research, conducted by the Bureau of Economic Geology and the Center for Energy Studies, includes the following areas of interest; geological studies depicting pressure gradients and thermal gradients, sand distribution and fault patterns, all of which are used in petroleum exploration; geophysical data for interpretation of seismic velocities based upon lithologic changes and subsurface discontinuities; sandstone consolidation data involving changes of permeabilities with depth and diagenetic histories of Cenozoic rocks in the Gulf Coast Basin--this work also covers fluid migration pathways and resulting rock-water interactions and has led to a better understanding of generation, maturation and accumulation of hydrocarbons; work on salinity of formation waters covering several areas of study, such as chemical analysis to anticipate scale and corrosion problems, and investigations of logging techniques to better ascertain salinity of use of well logs; reservoir continuity studies, together with computational modeling to assist in estimation of ultimate recoveries and formation drives; rock mechanics studies, which have recently led to the development of new models to account for creep and determine compressibilities of sandstones and shales in geopressured environments; co-production of gas and water in watered-out gas reservoirs

<i>NOTCH1</i> mutations in CLL.

<p>A) Position of <i>NOTCH1</i> mutations, superimposed on a cartoon showing the NOTCH1 protein. NRR, negative regulatory region; RAM, RAM domain; ANK, ankyrin repeat domain; TAD, transcriptional activation domain; PEST, degron domain. B) Representative confirmatory pyrosequencing results in CLL cases positive by deep sequencing for the <i>NOTCH1</i> codon 2514 del(CT) mutation. Results for a case with wild type codon 2514 sequences is shown in the upper panel. The lower two panels are cases in which deep sequencing revealed high (54.1%) or low numbers of mutated reads (3.3%). Reverse-sequences of the wild type (WT) and mutant (MUT) del(CT) alleles are shown for reference. “E” corresponds to the addition of enzyme to the reaction chamber, while “S” corresponds to the addition of the substrate. In this sequencing by synthesis assay, nucleotides are added to the reaction chamber sequentially in the order shown. Nucleotide incorporation releases pyrophosphate, which catalyses a reaction in which the number of photons produced is proportional to the number of nucleotides incorporated. The codon 2514 del(CT) mutation causes the appearance of a new signal corresponding to the incorporation of 3 G residues.</p

Relationship of NICD1 staining, <i>NOTCH1</i> genotype, and karyotype in CLL.

<p>A) Correlation between the fraction of mutated <i>NOTCH1</i> exon 34 reads and NICD1 staining. B) NICD1 staining in CLLs with and without mutated <i>NOTCH1</i> alleles. Filled squares correspond to CLLs with the NOTCH1 codon 2514 del(CT) mutation; black squares are tumors with a “high” fraction (>20%) of mutated reads, while gray squares are tumors with a “low” fraction (<5%) of mutated reads. The open square corresponds to a CLL with a nonsense mutation in codon 2444. C) Relationship of NICD1 staining to CLL karyotype. Open squares correspond to tumors with <i>NOTCH1</i> mutations.</p

NICD1 staining in normal formalin FPE human tissues.

<p>A) Oropharyngeal squamous mucosa. B) Skin. C, D) Thymus.</p