Vascular Signature Research Articles

Abstract 3828: An 18-gene signature for vascular invasion is associated with aggressive features and reduced survival in breast cancer

Abstract Introduction: Vascular invasion, i.e. tumor cells entering the vascular system, is considered to be an early step in the metastatic process and important for the progress of malignant tumors. By microarray and qPCR, we previously constructed an 18-gene signature in relation to vascular invasion among endometrial cancer patients. Here, we wanted to investigate the vascular invasion signature (VIS) among breast cancers patients. Methods: The 18-gene signature was studied in 11 gene expression data sets of breast cancer including information on 2423 patients. The genes in the signature were investigated in the external data sets and related to clinical data and follow-up information. We also wanted to examine if our signature was associated with other published signatures related to tumor progression. Results and Conclusions: High VIS showed associations with aggressive features of breast cancer such as high tumor grade, hormone receptor negativity, HER2 positive tumors, presence of a basal-like phenotype, reduced patient survival and response to cytostatic treatment. Correlations between VIS and other signatures for tumor progression indicate an involvement in similar biological processes. In conclusion, the vascular invasion signature showed strong and consistent associations with aggressive features of breast cancer. Citation Format: Monica Mannelqvist, Elisabeth Wik, Ingunn M. Stefansson, Lars A. Akslen. An 18-gene signature for vascular invasion is associated with aggressive features and reduced survival in breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3828. doi:10.1158/1538-7445.AM2014-3828

Abstract A027: Endothelial-like properties of triple-negative claudin-low breast cancer cells promote tumor-vascular interactions and tumor permeability

Abstract The different intrinsic subtypes of human breast tumors vary in rate and location of metastatic dissemination. In this study, we aimed to determine if tumor-associated vascular properties could help to explain the differences observed in metastagenicity. We used endothelial and breast cancer cell line models to develop gene expression signatures that measured vascular quantity and vascular proliferation. We tested these signatures, amongst other published endothelial signatures, and found that the genomic programs that measured vascular quantity, vascular proliferation, and a VEGF/Hypoxia-signature were the most highly expressed in claudin-low and basal-like tumors. We then tested the vascular signatures on five human breast cancer datasets (n>3,000) and identified two vascular signatures that added prognostic information to intrinsic subtype classification. Next, microvessel density (MVD) scores were obtained from a set of 70 tumors which were also subjected to gene expression profiling. Interestingly, several of the vascular signatures added metastasis-predicting information in addition to MVD (p<0.05). Similar to the tumor data, claudin-low cell lines also expressed high levels of vascular gene signatures as compared to luminal cell lines. Interestingly, pure claudin-low cell lines, and subsets of claudin-low-like cells within established basal-like cell cancer cell lines, exhibited endothelial-like morphology when culture on Matrigel. In vivo xenografts of claudin-low tumors, but not luminal tumors, extensively perfused injected contrast agent through paracellular spaces and non-vascular tumor-lined channels. Taken together, the endothelial-like characteristics of the cancer cells, combined with both the amount and the physiologic state of the vasculature, contribute to breast cancer metastatic progression. We hypothesize that the genetic signatures we have identified highlight patients that should respond most favorably to anti-vascular agents. Citation Format: J. Chuck Harrell, Adam Pfefferle, Nicole Zalles, Aleix Prat, Cheng Fan, Andrey Khramtsov, Olufunmilayo Olopade, Melissa Troester, Andrew Dudley, Charles Perou. Endothelial-like properties of triple-negative claudin-low breast cancer cells promote tumor-vascular interactions and tumor permeability. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research: Genetics, Biology, and Clinical Applications; Oct 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2013;11(10 Suppl):Abstract nr A027.

Defining vascular signatures of benign hepatic masses: role of MDCT with 3D rendering

Multidetector CT (MDCT) provides new opportunities for hepatic tumor characterization. By coupling high-resolution isotropic datasets with advanced post-processing tools, maps of tumor vascularity can be generated to elucidate characteristic findings. This two-part review describes a range of benign and malignant liver masses, with emphasis on IV contrast-enhanced MDCT features and vascular signatures that can be identified on 3D vascular mapping.

Gene-expression profiling of microdissected breast cancer microvasculature identifies distinct tumor vascular subtypes

IntroductionAngiogenesis represents a potential therapeutic target in breast cancer. However, responses to targeted antiangiogenic therapies have been reported to vary among patients. This suggests that the tumor vasculature may be heterogeneous and that an appropriate choice of treatment would require an understanding of these differences.MethodsTo investigate whether and how the breast tumor vasculature varies between individuals, we isolated tumor-associated and matched normal vasculature from 17 breast carcinomas by laser-capture microdissection, and generated gene-expression profiles. Because microvessel density has previously been associated with disease course, tumors with low (n = 9) or high (n = 8) microvessel density were selected for analysis to maximize heterogeneity for this feature.ResultsWe identified differences between tumor and normal vasculature, and we describe two subtypes present within tumor vasculature. These subtypes exhibit distinct gene-expression signatures that reflect features including hallmarks of vessel maturity. Potential therapeutic targets (MET, ITGAV, and PDGFRβ) are differentially expressed between subtypes. Taking these subtypes into account has allowed us to derive a vascular signature associated with disease outcome.ConclusionsOur results further support a role for tumor microvasculature in determining disease progression. Overall, this study provides a deeper molecular understanding of the heterogeneity existing within the breast tumor vasculature and opens new avenues toward the improved design and targeting of antiangiogenic therapies.

Defining vascular signatures of malignant hepatic masses: role of MDCT with 3D rendering

Malignant hepatic masses, both primary and metastatic lesions, have characteristic CT appearances and enhancement patterns. Owing to advances in CT resolution, high-quality vascular maps can be generated with 3D rendering tools to aid hepatic mass evaluation. These renderings enable identification of neovascularity, which is critical for distinguishing malignant from benign lesions, and facilitate identification of small hyperenhancing malignant hepatic tumors. In this review, CT features of malignant hepatic masses are discussed in conjunction with a demonstration of the role for 3D vascular mapping.

The dorsal sacral rami and branches: Sonographic visualisation of their vascular signature

ObjectivesThe differentiation of potential pain generating structures in the sacro-iliac region remains a diagnostic challenge. The aim of this proof of concept study was to evaluate the feasibility of using Doppler ultrasound to image the vascular components of dorsal sacral rami and their associated medial and lateral branches. Using the vascular signature of the dorsal sacral arteries to identify the dorsal sacral rami could assist with the diagnosis and treatment of localisable, extra-articular, posterior sacro-iliac joint pain. MethodsThe posterior sacral region was scanned in 30 healthy adults using an Acuson X300 ultrasound machine with 7.5 MHz high-resolution linear transducer in spectral Doppler mode to assess the feasibility of identifying and measuring the Resistive Index (RI) of the dorsal sacral arteries. ResultsThe vascular signature of the dorsal sacral rami and associated branches was identified in 62% of participants; S2 70%, S1 57% and S3 59%. More than two vascular signatures were recorded in 23 (76.7%) cases; in only two (6.67%) were no vascular signatures detected. Mean RI was 0.82 ± 0.11, increasing significantly (P < 0.05) at each caudal sacral level. No statistically significant difference in RI was observed between rami arteries and their branches or between genders. ConclusionsDoppler ultrasound may be useful in identifying the individual dorsal sacral rami (S1-3) and their associated branches. This may assist with selective imaging of the dorsal sacral rami and their branches in the diagnosis and treatment of chronic localisable pain in the posterior sacro-iliac region and facilitate investigation of the hypothesis of entrapment neuropathy in this region.

Molecular signatures of disease brain endothelia provide new sites for CNS-directed enzyme therapy

The brain vasculature forms an immense network such that most neural cells are in contact with a microvessel. Here we tested the hypothesis that endothelia lining these vessels can be harnessed to create a cellular reservoir of enzyme replacement therapy to diseased brain. As a model system, we used animals with central nervous system (CNS) deficits due to lysosomal storage disease (LSD). The basic premise is that recombinant enzyme expressed in, and secreted from, the vascular endothelia will be endocytosed by underlying neurons and glia, decreasing neuropathology. We screened a phage library in vivo to identify peptides that bound the vascular endothelia1 in diseased and wildtype mice. Surprisingly, epitopes binding diseased brain were distinct from those panned from normal brain. Moreover, different epitopes were panned out of different disease models, implying a unique vascular signature imparted by the disease state. Importantly, presentation of these epitopes on the capsid of adeno-associated virus (AAV) expanded the biodistribution of IV-injected AAV from predominantly liver to include the CNS. Peripheral injection of the epitope-modified AAVs expressing the enzymes lacking in LSD mice reconstituted enzyme activity throughout the brain and improved disease phenotypes in two distinct models.

Cortical spreading ischaemia is a novel process involved in ischaemic damage in patients with aneurysmal subarachnoid haemorrhage.

The term cortical spreading depolarization (CSD) describes a wave of mass neuronal depolarization associated with net influx of cations and water. Clusters of prolonged CSDs were measured time-locked to progressive ischaemic damage in human cortex. CSD induces tone alterations in resistance vessels, causing either transient hyperperfusion (physiological haemodynamic response) in healthy tissue; or hypoperfusion [inverse haemodynamic response = cortical spreading ischaemia (CSI)] in tissue at risk for progressive damage, which has so far only been shown experimentally. Here, we performed a prospective, multicentre study in 13 patients with aneurysmal subarachnoid haemorrhage, using novel subdural opto-electrode technology for simultaneous laser-Doppler flowmetry (LDF) and direct current-electrocorticography, combined with measurements of tissue partial pressure of oxygen (ptiO2). Regional cerebral blood flow and electrocorticography were simultaneously recorded in 417 CSDs. Isolated CSDs occurred in 12 patients and were associated with either physiological, absent or inverse haemodynamic responses. Whereas the physiological haemodynamic response caused tissue hyperoxia, the inverse response led to tissue hypoxia. Clusters of prolonged CSDs were measured in five patients in close proximity to structural brain damage as assessed by neuroimaging. Clusters were associated with CSD-induced spreading hypoperfusions, which were significantly longer in duration (up to 144 min) than those of isolated CSDs. Thus, oxygen depletion caused by the inverse haemodynamic response may contribute to the establishment of clusters of prolonged CSDs and lesion progression. Combined electrocorticography and perfusion monitoring also revealed a characteristic vascular signature that might be used for non-invasive detection of CSD. Low-frequency vascular fluctuations (LF-VF) (f < 0.1 Hz), detectable by functional imaging methods, are determined by the brain's resting neuronal activity. CSD provides a depolarization block of the resting activity, recorded electrophysiologically as spreading depression of high-frequency-electrocorticography activity. Accordingly, we observed a spreading suppression of LF-VF, which accompanied spreading depression of high-frequency-electrocorticography activity, independently of whether CSD was associated with a physiological, absent or inverse haemodynamic response. Spreading suppressions of LF-VF thus allow the differentiation of progressive ischaemia and repair phases in a fashion similar to that shown previously for spreading depressions of high-frequency-electrocorticography activity. In conclusion, it is suggested that (i) CSI is a novel human disease mechanism associated with lesion development and a potential target for therapeutic intervention in stroke; and that (ii) prolonged spreading suppressions of LF-VF are a novel ‘functional marker’ for progressive ischaemia.

Hysteroscopic management of cornual ectopic pregnancy

A 32 year old woman attended the accident and emergency department with a two day complaint of vaginal bleeding and cramping lower abdominal pain, and a brown vaginal discharge since her last menstrual period which was 24 days prior to presentation. She had discontinued the combined oral contraceptive pill two months prior to her last menstrual period. Her urine pregnancy test (Guest Medical, UK) was positive. On abdominal examination, there was tenderness noted in the right iliac fossa. Vaginal examination revealed no cervical excitation but there was right adnexal tenderness. A transabdominal and transvaginal ultrasound scan (Diagnostic Sonar, multifrequency abdominal probe and 6.5 Hz transvaginal probe) revealed an empty uterus with an endometrial thickness of 11 mm. There was a right-sided ovarian cyst measuring 77 54 66 mm and a small amount of free fluid in the pouch of Douglas. Her serum h-hCG quantitative assay (Immulite 2000, immunoassay calibrated to WHO 3rd International Standard) at that point was 2830 IU/mL. A diagnostic laparoscopy was performed the following day to exclude an ectopic pregnancy. At laparoscopy, the finding of a right ovarian cyst was confirmed, with normal tubes, uterus and left ovary. A laparoscopic right ovarian cystectomy was performed as this was presumed to be the cause of her pain. A repeat h-hCG level the following day (48 hours after admission) was 3116 IU/mL. A repeat ultrasound scan two days later (four days after admission) showed what was presumed to be a viable intrauterine pregnancy, which was high within the uterine fundus. The possibility of a cornual pregnancy was raised. It was decided to rescan her in two weeks. A repeat ultrasound scan two weeks later revealed a left-sided cornual ectopic pregnancy, with a 25 mm gestational sac containing a 6 mm fetal pole. There was a large vascular signature from the surrounding decidual reaction, no fetal heart activity was seen and there was no free fluid in the pouch of Douglas. The serum h-hCG level at this stage was 15,208 IU/mL. Management options were discussed with the patient and she opted for surgical management. The following day, a laparoscopy and hysteroscopy were performed. The laparoscopy confirmed the diagnosis of a left-sided cornual ectopic pregnancy. A dilute solution of vasopressin (20 units in 20 mL of normal saline) was injected around the pregnancy. A hysteroscopy (5 mm, 30j forward angled scope) with normal saline showed a pregnancy beyond the left tubal ostium. The hysteroscope was advanced beyond the dilated left ostium and the sac was ruptured. The fetus was seen floating in the irrigation fluid. The hysteroscope was then withdrawn and a No. 6 flexible suction cannula (Rocket Medical, UK) inserted into the uterine cavity. Under transabdominal ultrasound guidance, the flexible suction cannula was advanced into the cornual gestation. Suction was then applied under direct laparoscopic control to ensure that the cannula did not perforate the uterine cornu. The suction evacuation yielded products of conception, which were sent for histology. There was minimal bleeding and the patient was kept in the hospital for observation. The serum h-hCG level fell to 1929 IU/mL by the third post-operative day. She recovered well and was discharged home. She was followed up in the Early Pregnancy Assessment Unit with weekly serum h-hCG levels, which fell gradually and became undetectable by five weeks following the operation. The histology confirmed products of conception.