The BET family of proteins targets moloney murine leukemia virus integration near transcription start sites.

A hallmark of retroviral replication is integration of the viral genome into host cell DNA. This characteristic makes retrovirus-based vectors attractive delivery vehicles for gene therapy. However, adverse events in gene therapeutic trials, caused by activation of proto-oncogenes due to murine leukemia virus (MLV)-derived vector integration, hamper their application. Here, we show that bromodomain and extraterminal (BET) proteins (BRD2, BRD3, and BRD4) and MLV integrase specifically interact and colocalize within the nucleus of the cell. Inhibition of the BET proteins' chromatin interaction via specific bromodomain inhibitors blocks MLV virus replication at the integration step. MLV integration site distribution parallels the chromatin binding profile of BET proteins, and expression of an artificial fusion protein of the BET integrase binding domain with the chromatin interaction domain of the lentiviral targeting factor LEDGF/p75 retargets MLV integration away from transcription start sites and into the body of actively transcribed genes, conforming to the HIV integration pattern. Together, these data validate BET proteins as MLV integration targeting factors.

Evaluation of breast tumor response to neoadjuvant chemotherapy with tomographic diffuse optical spectroscopy: case studies of tumor region-of-interest changes.

PURPOSE: To evaluate two methods of summarizing tomographic diffuse optical spectroscopic (DOS) data through region-of-interest (ROI) analysis to differentiate complete from incomplete responses in patients with locally advanced breast cancer undergoing neoadjuvant treatment and to estimate the standard deviations of these methods for power analysis of larger study designs in the future. MATERIALS AND METHODS: Subjects participating in the HIPAA-compliant imaging study, approved by the institutional review board, provided written informed consent and were compensated for their examination participation. Seven of 16 cases in women with complete study data were analyzed by using both fixed- and variable-size (full-width-at-half-maximum) ROI measures of the DOS total hemoglobin concentration (Hb(T)), blood oxygen saturation, water fraction, optical scattering amplitude, and scattering power in the ipsilateral and contralateral breasts. Postsurgical histopathologic analysis was used to categorize patients as having a complete or incomplete treatment response. RESULTS: Average normalized change in Hb(T) was the only DOS parameter to show significant differences (P < or = .05) in the pathologic complete response (pCR) and pathologic incomplete response (pIR) outcomes in seven patients. Mean values of the changes for fixed-size ROIs were -64.2% +/- 50.8 (standard deviation) and 16.9% +/- 38.2 for the pCR and pIR groups, respectively, and those for variable-size ROIs were -96.7% +/- 91.8, and 14.1% +/- 26.7 for the pCR and pIR groups, respectively. CONCLUSION: Tomographic DOS may provide findings predictive of therapeutic response, which could lead to superior individualized patient treatment. SUPPLEMENTAL MATERIAL: http://radiology.rsnajnls.org/cgi/content/full/2522081202/DC1.

The correlation of in vivo and ex vivo tissue dielectric properties to validate electromagnetic breast imaging: initial clinical experience.

Electromagnetic (EM) breast imaging provides low-cost, safe and potentially a more specific modality for cancer detection than conventional imaging systems. A primary difficulty in validating these EM imaging modalities is that the true dielectric property values of the particular breast being imaged are not readily available on an individual subject basis. Here, we describe our initial experience in seeking to correlate tomographic EM imaging studies with discrete point spectroscopy measurements of the dielectric properties of breast tissue. The protocol we have developed involves measurement of in vivo tissue properties during partial and full mastectomy procedures in the operating room (OR) followed by ex vivo tissue property recordings in the same locations in the excised tissue specimens in the pathology laboratory immediately after resection. We have successfully applied all of the elements of this validation protocol in a series of six women with cancer diagnoses. Conductivity and permittivity gauged from ex vivo samples over the frequency range 100 Hz-8.5 GHz are found to be similar to those reported in the literature. A decrease in both conductivity and permittivity is observed when these properties are gauged from ex vivo samples instead of in vivo. We present these results in addition to a case study demonstrating how discrete point spectroscopy measurements of the tissue can be correlated and used to validate EM imaging studies.

Measurement of pressure-displacement kinetics of hemoglobin in normal breast tissue with near-infrared spectral imaging.

Applying localized external displacement to the breast surface can change the interstitial fluid pressure such that regional transient microvascular changes occur in oxygenation and vascular volume. Imaging these dynamic responses over time, while different pressures are applied, could provide selective temporal contrast for cancer relative to the surrounding normal breast. In order to investigate this possibility in normal breast tissue, a near-infrared spectral tomography system was developed that can simultaneously acquire data at three wavelengths with a 15 s time resolution per scan. The system was tested first with heterogeneous blood phantoms. Changes in regional blood concentrations were found to be linearly related to recovered mean hemoglobin concentration (Hb(T)) values (R(2)=0.9). In a series of volunteer breast imaging exams, data from 17 asymptomatic subjects were acquired under increasing and decreasing breast compression. Calculations show that a 10 mm displacement applied to the breast results in surface pressures in the range of 0-55 kPa depending on breast density. The recovered human data indicate that Hb(T) was reduced under compression and the normalized change was significantly correlated to the applied pressure with a p value of 0.005. The maximum Hb(T) decreases in breast tissue were associated with body mass index (BMI), which is a surrogate indicator of breast density. No statistically valid correlations were found between the applied pressure and the changes in tissue oxygen saturation (S(t)O(2)) or water percentage (H(2)O) across the range of BMI values studied.

Digital breast tomosynthesis: initial experience in 98 women with abnormal digital screening mammography.

OBJECTIVE: The purpose of our study was to compare the image quality of tomosynthesis with that of conventional mammography and to estimate the recall rate of screening when tomosynthesis is used in addition to mammography. MATERIALS AND METHODS: Women with an abnormal screening mammography were recruited sequentially. Consenting women underwent tomosynthesis of the affected breast corresponding to the views obtained with diagnostic mammography. The study radiologist compared the image quality, including lesion conspicuity and feature analysis, of tomosynthesis with diagnostic film-screen mammography and assessed the need for recall when tomosynthesis was added to digital screening mammography. Screening recalls were considered unnecessary when tomosynthesis did not show a corresponding abnormality or allowed definitely benign lesion characterization. Fisher's exact test was used to determine the association of equivalence and recall status with mammographic finding type. RESULTS: There were 99 digital screening recalls in 98 women. The image quality of tomosynthesis was equivalent (n = 51) or superior (n = 37) to diagnostic mammography in 89% (88/99). Finding type was significantly (p < 0.001) associated with equivalence. Approximately half--52/99 (52%)--of the findings would not have been recalled when digital screening mammography was supplemented with tomosynthesis. When adjusting for confounding conditions, the recall reduction was 40% (37/92). The likelihood of recall was also dependent on finding type (p = 0.004). CONCLUSION: Subjectively, tomosynthesis has comparable or superior image quality to that of film-screen mammography in the diagnostic setting, and it has the potential to decrease the recall rate when used adjunctively with digital screening mammography.

Electromagnetic breast imaging: results of a pilot study in women with abnormal mammograms.

PURPOSE: To prospectively assess quantitatively the inherent contrast of electromagnetic (EM) properties that can be imaged by using available technology in women with abnormal findings at conventional breast imaging who underwent subsequent biopsy. MATERIALS AND METHODS: The protocol was HIPAA compliant and approved by the institutional review board. All participants provided informed consent. Fifty-three women with normal (Breast Imaging Reporting and Data System [BI-RADS] category 1) and ninety-seven women with abnormal (BI-RADS category 4 or 5) screening mammograms were imaged with three EM imaging methods: electrical impedance spectroscopy (EIS), microwave imaging spectroscopy (MIS), and near-infrared spectral tomography (NIR). A region-of-interest (ROI) analysis was used to assess the EM image properties for comparison of findings with conventional image findings and correlation with specific pathologic parameters for women with abnormal findings. Statistical analyses were conducted. RESULTS: One hundred fifty participants (age range, 35-81 years) were included. EM image property contrast ratios of 150%-200% were found in breast abnormality ROIs relative to the ipsilateral breast background. Analysis of variance demonstrated significant differences in ROI image summaries of mammographically normal versus abnormal breasts for EIS, across diagnostic groups for NIR, and for MIS (analysis restricted to lesions larger than 1 cm(3)). Receiver operating curve (ROC) analysis of the EM properties for cancers among subjects with BI-RADS category 4 or 5, compared with the EM properties for the subjects with normal breasts (BI-RADS category 1), yielded areas under the ROC curve ranging from 0.67 to 0.81. Pathologic correlations with mean vessel density, mean vessel area, and epithelium-to-stroma ratio suggest a biological origin of the EM image properties associated with disease. CONCLUSION: Results from EM breast examinations provide statistical evidence of a mean increase in image contrast of 150%-200% between abnormal (benign and malignant) and normal breast tissue.

Image-guided optical spectroscopy provides molecular-specific information in vivo: MRI-guided spectroscopy of breast cancer hemoglobin, water, and scatterer size.

A multimodality instrument that integrated optical or near-infrared spectroscopy into a magnetic resonance imaging (MRI) breast coil was used to perform a pilot study of image-guided spectroscopy on cancerous breast tissue. These results are believed to be the first multiwavelength spectroscopic images of breast cancer using MRI-guided constraints, and they show the cancer tumor to have high hemoglobin and water values, decreased oxygen saturation, and increased subcellular granularity. The use of frequency-domain diffuse tomography methods at many wavelengths provides the spectroscopy required for recovering maps of absorbers and scattering spectra, but the integration with MRI allows these data to be recovered on an image field that preserves high resolution and fuses the two data sets together. Integration of molecular spectroscopy into standard clinical MRI can be achieved with this approach to spectral tomography.

Initial clinical experience with microwave breast imaging in women with normal mammography.

RATIONALE AND OBJECTIVES: We have developed a microwave tomography system for experimental breast imaging. MATERIALS AND METHODS: In this article, we illustrate a strategy for optimizing the coupling liquid for the antenna array based on in vivo measurement data. We present representative phantom experiments to illustrate the imaging system's ability to recover accurate property distributions over the range of dielectric properties expected to be encountered clinically. To demonstrate clinical feasibility and assess the microwave properties of the normal breast in vivo, we summarize our initial experience with microwave breast exams of 43 women with negative mammography according to the Breast Imaging Reporting and Data System (BI-RADS 1). RESULTS: The clinical results show a high degree of bilateral symmetry in the whole breast average microwave properties. Focal assessments of microwave properties are associated with breast tissue composition evaluated through radiographic density categorization verified through magnetic resonance image correlation in selected cases. Specifically, both whole-breast average and local microwave properties increase with increasing radiographic density, in which the latter exhibits a more substantial rise. CONCLUSION: These findings support our hypothesis that water content variations in the breast play an influential role in dictating the overall dielectric property distributions and indicate that the microwave properties in the breast are more heterogeneous than previously believed based on ex vivo property measurements reported in the literature.

Image reconstruction of effective Mie scattering parameters of breast tissue in vivo with near-infrared tomography.

A method for image reconstruction of the effective size and number density of scattering particles is discussed within the context of interpreting near-infrared (NIR) tomography images of breast tissue. An approach to use Mie theory to estimate the effective scattering parameters is examined and applied, given some assumptions about the index of refraction change expected in lipid membrane-bound scatterers. When using a limited number of NIR wavelengths in the reduced scattering spectra, the parameter extraction technique is limited to representing a continuous distribution of scatterer sizes, which is modeled as a simple exponentially decreasing distribution function. In this paper, image formation of effective scatterer size and number density is presented based on the estimation method. The method was evaluated with Intralipid phantom studies to demonstrate particle size estimation to within 9% of the expected value. Then the method was used in NIR patient images, and it indicates that for a cancer tumor, the effective scatterer size is smaller than the background breast values and the effective number density is higher. In contrast, for benign tumor patients, there is not a significant difference in effective scatterer size or number density between tumor and normal tissues. The method was used to interpret magnetic resonance imaging-coupled NIR images of adipose and fibroglandular tissues, and it indicated that the fibroglandular tissue has smaller effective scatterer size and larger effective number density than the adipose tissue does.

In vivo hemoglobin and water concentrations, oxygen saturation, and scattering estimates from near-infrared breast tomography using spectral reconstruction.

RATIONALE AND OBJECTIVES: Near-infrared (NIR) imaging has its niche in quantifying and characterizing functional changes in tissue relating to vascularity and metabolic status. Here, NIR tomography was applied to study mammographically normal breast tissue in vivo by evaluating relationships between functional parameters so obtained to clinical representers in an effort to understand factors influencing tissue compositional changes. MATERIALS AND METHODS: A new spectral reconstruction method that is considered to provide the most accurate estimates of hemoglobin level, oxygen saturation, water fraction, scattering power, and amplitude was used to assess healthy breast tissue imaged in vivo by means of NIR tomography. The approach directly recovers functional parameters with inherent inclusion of spectral behavior enforced through the incorporation of a priori model assumptions. Sixty subjects were imaged by using a frequency-domain instrument followed by spectral image reconstruction and statistical analysis for significant correlations. RESULTS: The new analysis shows statistically significant inverse correlations between body mass index and breast total hemoglobin and water fractions. Water fraction also correlated inversely with age and separated certain categories of breast density. Average scatter power was indicative of breast radiographic density composition, whereas scatter amplitude varied inversely with breast diameter. Total hemoglobin correlated with water fraction, whereas water correlated with scatter power. CONCLUSION: The changes observed here are attributable to volume fraction alterations and provide some of the most comprehensive data on breast composition variations with demographic factors.

Near-infrared characterization of breast tumors in vivo using spectrally-constrained reconstruction.

Multi-wavelength Near-Infrared (NIR) Tomography was utilized in this study to non-invasively quantify physiological parameters of breast tumors using direct spectral reconstruction. Frequency domain NIR measurements were incorporated with a new spectrally constrained direct chromophore and scattering image reconstruction algorithm, which was validated in simulations and experimental phantoms. Images of total hemoglobin, oxygen saturation, water, and scatter parameters were obtained with higher accuracy than previously reported. Using this spectral approach, in vivo NIR images are presented and interpreted through a series of case studies (n=6 subjects) having differing abnormalities. The corresponding mammograms and ultrasound images are also evaluated. Three of six cases were malignant (infiltrating ductal carcinomas) and showed higher hemoglobin (34-86% increase), a reduction in oxygen saturation, an increase in water content as well as scatter changes relative to surrounding normal tissue. Three of six cases were benign, two of which were diagnosed with fibrocystic disease and showed a dominant contrast in water, consistent with fluid filled cysts. Scatter amplitude was the main source of contrast in the volunteer with the benign condition fibrosis, which typically contains denser collagen tissue. The changes monitored correspond to physiological changes associated with angiogenesis, hypoxia and cell proliferation anticipated in cancers. These changes represent potential diagnostic indicators, which can be assessed to characterize breast tumors.

Characterization of hemoglobin, water, and NIR scattering in breast tissue: analysis of intersubject variability and menstrual cycle changes.

Near-infrared imaging was used to quantify typical values of hemoglobin concentration, oxygen saturation, water fraction, scattering power, and scattering amplitude within the breast tissue of volunteer subjects. A systematic study of the menstrual variations in these parameters was carried out by measuring a group of seven premenopausal normal women (aged 41 to 47 years) in the follicular (days 7 to 14 of the cycle) and secretory phases (days 21 to 28) of the cycle, for two complete menstrual cycles. An average increase in hemoglobin concentration of 2.6 microM or 13% of the background breast values was observed in the secretory phase relative to the follicular phase (p<0.0001), but no other average near-infrared parameter changes were significant. While repeatable and systematic changes were observed in all parameters for individual subjects, large intersubject variations were present in all parameters. In a survey of thirty-nine normal subjects, the total hemoglobin varied from 9 to 45 microM, with a systematic correlation observed between total hemoglobin concentration and breast radiographic density. Scattering power and scattering amplitude were also correlated with radiographic density, but oxygen saturation and water fraction were not. Images of breast lesions indicate that total hemoglobin-based contrast can be up to 200% relative to the background in the same breast. Yet, since the background hemoglobin values vary considerably among breasts, the maximum hemoglobin concentrations observed in cancer tumors may vary considerably as well. In light of these observations, it may be important to use hemoglobin contrast values relative to the background for a given breast, rather than absolute hemoglobin contrast when trying to compare the features of breast lesions among subjects.

Multi-frequency electrical impedance tomography of the breast: new clinical results.

Electrical impedance tomography (EIT) has been used in the recent past for a number of clinical applications. In this work we present recent tomographic and spectroscopic findings for breast imaging from clinical exams completed at Dartmouth. The results presented here are based on 18 normal and 24 abnormal subjects. The participants were classified as normal or abnormal using the American College of Radiology (ACR) indexing system for mammograms. The EIT data were collected for ten discrete frequencies in the range 10 kHz-1 MHz using a single array of 16 electrodes. The finite element method was used to reconstruct the images. The images were examined visually and were compared with mammograms. The results were also analyzed based on zonal averages of property values and breast tissue radiodensities. Statistical analysis showed a significance difference between the mean conductivity and permittivity values of normal and abnormal subjects for various zones defined on the reconstructed images. Tissues with high radiodensity also had increased conductivity and permittivity.

Interpreting hemoglobin and water concentration, oxygen saturation, and scattering measured in vivo by near-infrared breast tomography.

Near-infrared spectroscopic tomography was used to measure the properties of 24 mammographically normal breasts to quantify whole-breast absorption and scattering spectra and to evaluate which tissue composition characteristics can be determined from these spectra. The absorption spectrum of breast tissue allows quantification of (i) total hemoglobin concentration, (ii) hemoglobin oxygen saturation, and (iii) water concentration, whereas the scattering spectrum provides information about the size and number density of cellular components and structural matrix elements. These property data were tested for correlation to demographic information, including subject age, body mass index, breast size, and radiographic density. Total hemoglobin concentration correlated inversely to body mass index, likely because lower body mass indicates proportionately less fat and more glandular tissue, and glandular tissue contains greater vascularity, hence, more total hemoglobin. Optical scattering was correlated to breast diameter, subject age, and radiographic density. In the radiographic density, fatty breasts had low scattering power and extremely dense breasts had higher values. This observation is consistent with low attenuation of conventional x-rays with fat and higher attenuation in glandular tissues. Optically, fatty tissues have large scatterers leading to a low scattering power, whereas glandular or fibrous tissues have more cellular and collagen-based structures that lead to high scattering power. The study presents correlative data supporting the hypothesis that optical measurements of absorption and scattering can provide physiologically relevant information about breast tissue composition. These breast constituents vary significantly between individuals and can be altered because of changes in breast physiology or pathological state.