Publications

2017

February, 17th
Filtration-based enrichment of circulating tumor cells from all prostate cancer risk groups.

Filtration-based enrichment of circulating tumor cells from all prostate cancer risk groups.

  • 1Manitoba Institute of Cell Biology, University of Manitoba, CancerCare Manitoba, 675 McDermot Ave, Winnipeg, Manitoba, Canada MB R3E 0V9; Systems Biology Research Centre, School of Life Sciences, University of Skövde, Skövde, Sweden; Department of Clinical Genetics, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
  • 2Department of Surgery, Manitoba Prostate Center, University of Manitoba, Winnipeg, Manitoba, Canada.
  • 3Manitoba Institute of Cell Biology, University of Manitoba, CancerCare Manitoba, 675 McDermot Ave, Winnipeg, Manitoba, Canada MB R3E 0V9. Electronic address: Sabine.Mai@umanitoba.ca.

Abstract

OBJECTIVE:

To combine circulating tumor cell (CTC) isolation by filtration and immunohistochemistry to investigate the presence of CTCs in low, intermediate, and high-risk prostate cancer (PCa). CTCs isolated from these risk groups stained positive for both cytokeratin and androgen receptors, but negative for CD45.

PATIENTS AND METHODS:

Blood samples from 41 biopsy confirmed patients with PCa at different clinical stages such as low, intermediate, and high risk were analyzed. The samples were processed with the ScreenCell filtration device and PCa CTCs were captured for all patients. The isolated CTCs were confirmed PCa CTCs by the presence of androgen receptors and cytokeratins 8, 18, and 19 that occurred in the absence of CD45 positivity. PCa CTC nuclear sizes were measured using the TeloView program.

RESULTS:

The filtration-based isolation method used permitted the measurement of the average nuclear size of the captured CTCs. CTCs were identified by immunohistochemistry in low, intermediate, and high-risk groups of patients with PCa.

CONCLUSION:

CTCs may be found in all stages of PCa. These CTCs can be used to determine the level of genomic instability at any stage of PCa; this will, in the future, enable personalized patient management.

Adebayo J Urological Oncology 2017

January, 6th
Detection of Circulating Tumour Cells and Survival of Patients with Non-small Cell Lung Cancer.

Detection of Circulating Tumour Cells and Survival of Patients with Non-small Cell Lung Cancer.

  • 1Department of Thoracic Surgery, Royal Brompton and Harefield NHS Foundation Trust, London, U.K.
  • 2Department of Histopathology, Royal Brompton and Harefield NHS Foundation Trust, London, U.K.
  • 3Department of Thoracic Surgery, Royal Brompton and Harefield NHS Foundation Trust, London, U.K. v.anikin@rbht.nhs.uk.

Abstract

BACKGROUND:

Detection of circulating tumour cells (CTCs) in the peripheral blood of lung cancer patients may predict survival. Various platforms exist that allow capture of these cells for further analysis; little work however, has been done with the ScreenCell device, an antibody-independent CTC platform. The aim of our study was to evaluate the ScreenCell device for detection of CTCs in lung cancer patients and to establish correlations of these findings with survival.

MATERIALS AND METHODS:

Twenty-three patients, nine males, and fourteen females, underwent surgical treatment from February to May 2014 for non-small cell lung cancer. Thirteen patients had adenocarcinoma and ten squamous cell carcinoma, while eight were at an early stage (I-II) and five at a later stage (III-IV). Blood samples were obtained prior to surgery and following filtration through the ScreenCell device, were independently reviewed by 2 consultant pathologists.

RESULTS:

The pathologists were able to independently identify CTCs in 78.3% (N=18) and 73.9% (N=17) of the cases examined, with overall 80.6% in early stages compared to 60.0% in late stages. The median survival times of positive vs. negative for CTC patients were 1011 and 711 days respectively, with a survival percentage rate of 77.8% and 60% in positive and negative CTC cohorts respectively.

CONCLUSION:

The results of this study suggest that the presence of CTCs analyzed by ScreenCell did not necessarily lead to a poorer prognosis in patients with lung cancer after curative surgery.

Detection of Circulating Tumour Cells and Survival of Patients with Non-small Cell Lung Cancer

2016

October, 3rd
Detection and Characterization of Circulating Tumor Associated Cells in Metastatic Breast Cancer

Article
Detection and Characterization of Circulating Tumor Associated Cells in Metastatic Breast Cancer
Zhaomei Mu 1,*, Naoual Benali-Furet 2, Georges Uzan 2, Anaëlle Znaty 2, Zhong Ye 3,
Carmela Paolillo 4, Chun Wang 3, Laura Austin 3, Giovanna Rossi 1, Paolo Fortina 4,5,
Hushan Yang 3 and Massimo Cristofanilli 1,*
1 Department of Medicine-Hematology and Oncology, Robert H Lurie Comprehensive Cancer Center,
Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA; giovirossi85@yahoo.it
2 ScreenCell SA, Sarcelles 95200, France; benali@screencell.com (N.B.-F.); guzan@screencell.com (G.U.);
aznaty@screencell.com (A.Z.)
3 Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University,
Philadelphia, PA 19107, USA; Zhong.Ye@jefferson.edu (Z.Y.); Chun.Wang@jefferson.edu (C.W.);
laustin@gmail.com (L.A.); hushan.yang@jefferson.edu (H.Y.)
4 Department of Cancer Biology, Sidney Kimmel Cancer Center, Thomas Jefferson University,
Philadelphia, PA 19107, USA; carmela.Paolillo@jefferson.edu (C.P.); paolo.Fortina@jefferson.edu (P.F.)
5 Department of Molecular Medicine, University of Rome “Sapienza”, Rome 00185, Italy
* Correspondence: zhaomei.mu@northwestern.edu (Z.M.); massimo.cristofanilli@nm.org (M.C.);
Tel.: +1-312-503-5489 (Z.M.); +1-312-503-5488 (M.C.)
Academic Editor: Dario Marchetti
Received: 5 August 2016; Accepted: 23 September 2016; Published: 30 September 2016
Abstract: The availability of blood-based diagnostic testing using a non-invasive technique holds
promise for real-time monitoring of disease progression and treatment selection. Circulating tumor
cells (CTCs) have been used as a prognostic biomarker for the metastatic breast cancer (MBC).
The molecular characterization of CTCs is fundamental to the phenotypic identification of malignant
cells and description of the relevant genetic alterations that may change according to disease
progression and therapy resistance. However, the molecular characterization of CTCs remains
a challenge because of the rarity and heterogeneity of CTCs and technological difficulties in the
enrichment, isolation and molecular characterization of CTCs. In this pilot study, we evaluated
circulating tumor associated cells in one blood draw by size exclusion technology and cytological
analysis. Among 30 prospectively enrolled MBC patients, CTCs, circulating tumor cell clusters (CTC
clusters), CTCs of epithelial–mesenchymal transition (EMT) and cancer associated macrophage-like
cells (CAMLs) were detected and analyzed. For molecular characterization of CTCs, size-exclusion
method for CTC enrichment was tested in combination with DEPArray™ technology, which allows
the recovery of single CTCs or pools of CTCs as a pure CTC sample for mutation analysis. Genomic
mutations of TP53 and ESR1 were analyzed by targeted sequencing on isolated 7 CTCs from a patient
with MBC. The results of genomic analysis showed heterozygous TP53 R248W mutation from one
single CTC and pools of three CTCs, and homozygous TP53 R248W mutation from one single CTC and
pools of two CTCs. Wild-type ESR1 was detected in the same isolated CTCs. The results of this study
reveal that size-exclusion method can be used to enrich and identify circulating tumor associated
cells, and enriched CTCs were characterized for genetic alterations in MBC patients, respectively.
Keywords: metastatic breast cancer (MBC); circulating tumor associated cells; circulating tumor
cells (CTCs); circulating tumor cell clusters (CTC clusters); epithelial–mesenchymal transition (EMT);
cancer associated macrophage-like cells (CAMLs); size-exclusion technology

detection-and-characterization-of-circulating-tumor-associated-cells-in-metastatic-breast-cancer-2016

May, 30th
Droplet Digital PCR of circulating tumor cells from colorectal cancer patients can predict KRAS mutations before surgery

Droplet Digital PCR of circulating tumor cells from colorectal cancer patients can predict KRAS mutations before surgery.
Jérôme Alexandre Denis1,2,3, Alexia Patroni4, Erell Guillerm1,5, Dominique Pépin2, Naoual Benali-Furet6, Janine Wechsler6, Gilles Manceau1,4, Maguy Bernard2, Florence Coulet1,5; Annette K. Larsen3, Mehdi Karoui1,4, Jean-Marc Lacorte1,2, 7
1. Sorbonne Universités, UPMC Univ. Paris 06, F-75005, Paris, France.
2. Assistance Publique-Hôpitaux de Paris, Pitié-Salpêtrière Hospital, Department of Oncology and Endocrine Biochemistry, Paris, France.
3. Cancer Biology and Therapeutics, Centre de Recherche Saint-Antoine, Institut National de la Santé et de la Recherche Médicale (INSERM) U938 and Institut Universitaire de Cancérologie (IUC), Université Pierre et Marie Curie (UPMC), Sorbonne Universities, Paris, France.
4. Assistance Publique-Hôpitaux de Paris, Pitié-Salpêtrière Hospital, Department of Digestive and Hepato-Pancreato-Biliary Surgery, Paris, France.
5. Assistance Publique-Hôpitaux de Paris, Pitié-Salpêtrière Hospital, Department of oncogenetics and molecular angiogenetics, Paris, France.
6. ScreenCell SA, Sarcelles, France
7. INSERM, UMR_S 1166, Institute of cardiometabolism and nutrition. ICAN. Paris, France

Abstract:
In colorectal cancer (CRC), KRAS mutations are a strong negative predictor for treatment with the EGFR-targeted antibodies cetuximab and panitumumab. Since it can be difficult to obtain appropriate tumor tissues for KRAS genotyping, alternative methods are required. Circulating tumor cells (CTCs) are believed to be representative of the tumor in real time. In this study, we explored the capacity of a size-based device for capturing CTCs coupled with a multiplex KRAS screening assay using droplet digital PCR (ddPCR). We showed that it is possible to detect a mutant ratio of 0.05 % and less than one KRAS mutant cell per mL total blood with ddPCR compared to about 0.5% and 50-75 cells for TaqMeltPCR and HRM. Next, CTCs were isolated from the blood of 35 patients with CRC at various stage of the disease. KRAS genotyping was successful for 86% (30/35) of samples with a KRAS codon 12/13 mutant ratio of 57% (17/30). In contrast, only one patient was identified as KRAS mutant when size-based isolation was combined with HRM or TaqMeltPCR. KRAS status was then determined for the 26 available formalin-fixed paraffin-embedded tumors using standard procedures. The concordance between the CTCs and the corresponding tumor tissues was 77% with a sensitivity of 83%. Taken together, the data presented here suggest that is feasible to detect KRAS mutations in CTCs from blood samples of CRC patients which are predictive for those found in the tumor. The minimal invasive nature of this procedure in combination with the high sensitivity of ddPCR might provide in the future an opportunity to monitor patients throughout the course of disease on multiple levels including early detection, prognosis, treatment and relapse as well as to obtain mechanistic insight with respect to tumor invasion and metastasis.

The paper has been accepted for publication in Molecular Oncology, in press.

2015

December, 10th
SABCs 2015: Detection and Characterization of CTCs Isolated by ScreenCell® Size Exclusion Technology in Metastatic Breast Cancer

SABCs 2015 : Poster Session 2 – Thursday, December 10 7:30 am – 9:00 am

https://www.sabcs.org/Program/Poster-Sessions/Poster-Session-2

Background: Circulating Tumor cells (CTCs) detection has prognostic and predictive implications in patients with metastatic breast cancer (MBC). Genomic and phenotypic analysis of CTCs hold enormous promise as blood-based molecular characterization and monitoring disease progression and treatment benefit with a strong potential to be translated into more individualized targeted treatments. FDA-approved CellSearch™ detection allows only enumeration of CTCs expressing EpCAM without molecular characterization. CTCs represent very heterogeneous populations of tumorigenic cancer cells and some subpopulations have undergone epithelial-Mesenchymal transition (EMT), which is associated metastasis process and an unfavourable outcome. EpCAM-based enrichment technique has failed to detect EMT subpopulations due to the decreased expression or loss of epithelial markers. Non-EpCAM-based approaches are needed for identifying EMT CTCs. The ScreenCell® devices are single-use and low-cost innovative devices that use a filter for enrichment-free isolation of CTCs by a two-steps combining size-based separation and staining using different markers. The DEPArray™ system is the ideal downstream isolation system to collect single or pooled CTCs for molecular and genetic analysis. In this study, we evaluated the feasibility of achieving CTCs detection/enumeration using ScreenCell® filtration followed by single cell isolation with the DEPArray™ in MBC patients.

Methods: The first part of the study consisted in evaluating CTCs detection/enumeration in 30 patients with stage III and stage IV breast cancer. 3 mL of whole blood in an EDTA or Transfix tubes was collected and processed on the ScreenCell® Cyto device following the instructions of the supplier. CTCs were stained with cytokeratin (CK-8, 18, and 19), leukocyte antigen (CD45), and a nuclear dye (DAPI) and counted under fluorescence microscope. CTCs were identified as positive staining for CK and DAPI and negative staining for CD45 (CK+/DAPI+CD45-). In the second part, After enrichment, CTCs were stained with CK, CD45, and DAPI and sorted with DEPArray™ Platform (Silicon Biosystems, Inc). Single CTCs were collected and the DNA of each single CTCs was amplified with Ampli1™ WGA kit, and the genome integrity index (GII) was assessed by Ampli1™ QC kit (Silicon Biosystems, Inc). Library was constructed and whole exome sequencing (WES) of DNA mutations was conducted.

Results: Twenty patient samples had CTCs detected (66.7%), the number of CTCs was 1 to 347 per 3.0 ml of whole blood. CTC-clusters were detected in 7 patient samples (23.3%). Single CTCs were collected on DEPArray™ platform after enrichment with ScreenCell filtration. GII was confirmed with the presence of short, medium, and long DNA fragments (3 to 4 PCR bands) in the WGA library by PCR-based assay. All collected CTCs showed high GII as measured by Ampli1™ QC kit (GII ≥ 3) for WES of DNA mutations. The data analysis of WES results is under processing.

Conclusions: ScreenCell® filtration is simple and effective devices to isolate CTCs and identify CTC-clusters. Isolation of single cells for molecular analysis using the combination of ScreenCell® filtration and DEPArray™ Platform is feasible for genetic characterization of CTCs.

Authors: Zhaomei Mu1, Naoual Benali-Furet2, Georges Uzan2, Zhong Ye1, Carmela Paolillo1, Laura Austin1, Chun Wang1, Rebecca Jaslow1, Hushan Yang1, Paolo Fortina1,
Massimo Cristofanilli1

Institutions: 1Department of Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, United States, 19107 and 2ScreenCell, Sarcelles, France, 95200.

SABCs-Poster P2 02 14 Massimo Cristofanilli dec 2015

October, 9th
A comparative analysis of cancer hotspot mutation profiles in circulating tumour cells, circulating tumour DNA and matched primary lung tumour

A comparative analysis of cancer hotspot mutation profiles in circulating tumour cells, circulating tumour DNA and matched primary lung tumour
Maria Leung2, Maxim Freidin1, Dasha Freidina1, Sanjay Popat3, Andrew Nicholson2, Alexandra Rice2, Angeles Montero Fernandez2, Eric Lim1,2
1National Heart and Lung Institute, Imperial College London; 2Royal Brompton Hospital, London; 3Royal Marsden Hospital, London

The aim of our work is to report the concordances between CTCs and ctDNA versus the primary FFPE tumour mutations using a NGS hotspot panel.

“Our results suggest on a next generation sequencing platform that the global genetic variant profile between DNA extracted from CTC had good agreement with FFPE primary tumour tissue, and the agreement between ctDNA and FFPE was much poorer!” said consultant thoracic surgeon Dr Eric Lim from Royal Brompton Hospital, London.

CTC vs cfDNA Lung Cancer

July, 30th
Prevalence and number of circulating tumour cells and microemboli at diagnosis of advanced NSCLC.

Prevalence and number of circulating tumour cells and microemboli at diagnosis of advanced NSCLC.

Abstract

PURPOSE:

Timing and magnitude of blood release of circulating tumour cells (CTC) and circulating tumour microemboli (CTM) from primary solid cancers are uncertain. We investigated prevalence and number of CTC and CTM at diagnosis of advanced non-small cell lung cancer (NSCLC).

METHODS:

Twenty-eight consecutive patients with suspected stage III-IV lung cancer gave consent to provide 15 mL of peripheral blood soon before diagnostic CT-guided fine-needle aspiration biopsy (FNAB). CTC and CTM (clusters of ≥3 CTC) were isolated by cell size filtration (ScreenCell), identified and counted by cytopathologists using morphometric criteria and (in 6 cases) immunostained for vimentin.

RESULTS:

FNAB demonstrated NSCLC in 26 cases. At least one CTC/3 mL blood (mean 6.8 ± 3.7) was detected in 17 (65 %) and one CTM (mean 4.5 ± 3.3) in 15 (58 %) of 26 NSCLC cases. No correlation between number of CTC or CTM and tumour type or stage was observed. Neoplastic cells from both FNA and CTC/CTM were positive for vimentin but heterogeneously.

CONCLUSIONS:

CTC can be detected in two-thirds and CTM in more than half of patients with advanced NSCLC at diagnosis. Reasons underlying lack of CTC and CTM in some advanced lung cancers deserve further investigations.

Prevalence and number of circulating tumour cells

July, 2nd
Circulating Tumor Cells in Diagnosing Lung Cancer: Clinical and Morphologic Analysis.

Circulating Tumor Cells in Diagnosing Lung Cancer: Clinical and Morphologic Analysis.

Fiorelli A, Accardo M, Carelli E, Angioletti D, Santini M, Di Domenico M.

Ann Thorac Surg. 2015 Jun;99(6):1899-905. doi: 10.1016/j.athoracsur.2014.11.049. Epub 2015 Feb 10.

Abstract

BACKGROUND:

The purpose of this study was to evaluate the value of circulating non-hematologic cells to differentiate benign from malignant lung lesions and their comparison with clinico-histologic features of corresponding primary lesions.

METHODS:

Circulating cells were isolated by size method from peripheral blood of 77 patients with malignant (n = 60) and benign (n = 17) lung lesions. They were morphologically classified as cells with malignant feature; cells with uncertain malignant feature; and cells with benign feature; then statistically correlated with clinico-cytopathologic characteristics of corresponding lung lesion.

RESULTS:

Malignant circulating cells were detected in 54 of 60 (90%) malignant patients, and in 1 of 17 (5%) benign patients; benign circulating cells in 1 of 60 (1%) malignant patients and in 15 of 17 (88%) benign patients; and circulating cells with uncertain malignant aspect in 5 of 60 (8%) malignant patients and 1 of 17 (5%) benign patients. For a malignant circulating cells count greater than 25, sensitivity and specificity were 89% and 100%, respectively. The count was significantly correlated with stage, size, and standard uptake value of primary tumor. In 39 of 54 (72%) cases, the malignant circulating cells allowed a specific histologic diagnosis of the corresponding primary tumor after immunohistochemical analysis.

CONCLUSIONS:

Malignant circulating cells may be a valid marker in the diagnostic workup of lung lesions. However, our resuts should be corroborated by larger future studies especially for patients having small nodules.

Circulating Tumor Cells in Diagnosing Lung Cancer Clinical and Morphologic Analysis

July, 2nd
Circulating Tumour Cells in Patients with Malignant Lung Tumors Undergoing Radio-frequency Ablation

Circulating Tumour Cells in Patients with Malignant Lung Tumors Undergoing Radio-frequency Ablation.

Anticancer Res. 2015 May;35(5):2823-6.

Abstract

BACKGROUND/AIM:

Radiofrequency ablation (RFA) is an increasingly utilised technique in patients with surgically-untreatable lesions. The effect of this therapy on circulating tumor cells (CTCs) is unknown. As far as we are aware of, this is the first study to evaluate the effects of RFA on CTCs in patients with malignant lung tumors immediately post-treatment.

PATIENTS AND METHODS:

Nine patients with primary or metastatic lung tumors underwent RFA therapy from June to November 2013. Blood samples were taken before and after RFA, and filtered through the ScreenCell CTC capture device.

RESULTS:

A general increase in CTCs in 7 out of the 9 cases was found, the largest increases were seen in the metastatic group.

CONCLUSION:

This study demonstrates that the manipulation and ablative procedure of lung tumors leads to immediate dissemination of tumor cells, the effects of which are unknown and require further investigation.

July, 2nd
Circulating tumour cells in patients with lung cancer undergoing endobronchial cryotherapy.

Circulating tumour cells in patients with lung cancer undergoing endobronchial cryotherapy.

Cryobiology. 2015 Jun 2. pii: S0011-2240(15)00175-3. doi: 10.1016/j.cryobiol.2015.06.001. [Epub ahead of print]

Abstract

Early diagnosis of lung cancer still poses a major issue, with a large proportion of patients diagnosed at late stages. Therapeutic options and treatment remain limited in these patients. In most cases only palliative therapies are available to alleviate any severe symptoms. Endobronchial cryotherapy (EC) is one form of palliative treatment offered to patients with obstructive airway tumours. Although successful, the impact on circulating tumour cell (CTCs) spread has not been investigated in detail. This study recruited 20 patients awaiting EC treatment. Baseline and post EC blood samples were analysed for presence of CTCs. Results showed an increase in CTCs following EC in 75% of patients. Significant increases were noticeable in some cases. Although EC is a well-accepted modality of treatment to alleviate symptoms, it may lead to an increase in CTCs, which in turn may have implications for tumour dissemination and metastatic spread.

Circulating tumour cells in patients with lung cancer undergoing endobronchial cryotherapy.

April, 2nd
Circulating Tumor Cells Found in Patients With Localized and Advanced Pancreatic Cancer.

Pancreas. 2015 Mar 27. [Epub ahead of print]

Circulating Tumor Cells Found in Patients With Localized and Advanced Pancreatic Cancer.

Kulemann B1, Pitman MB, Liss AS, Valsangkar N, Fernández-Del Castillo C, Lillemoe KD, Hoeppner J, Mino-Kenudson M, Warshaw AL, Thayer SP.

Abstract

OBJECTIVES:

Isolation of circulating tumor cells (CTCs) holds the promise of diagnosing and molecular profiling cancers from a blood sample. Here, we test a simple new low-cost filtration device for CTC isolation in patients with pancreatic ductal adenocarcinoma (PDAC).

METHODS:

Peripheral blood samples drawn from healthy donors and PDAC patients were filtered using ScreenCell devices, designed to capture CTCs for cytologic and molecular analysis. Giemsa-stained specimens were evaluated by a pancreatic cytopathologist blinded to the histological diagnosis. Circulating tumor cell DNA was subjected to KRAS mutational analysis.

RESULTS:

Spiking experiments demonstrated a CTC capture efficiency as low as 2 cells/mL of blood. Circulating tumor cells were identified by either malignant cytology or presence of KRAS mutation in 73% of 11 patients (P = 0.001). Circulating tumor cells were identified in 3 of 4 patients with early (≤American Joint Committee on Cancer stage IIB) and in 5 of 7 patients with advanced (≥ American Joint Committee on Cancer stage III) PDAC. No CTCs were detected in blood from 9 health donors.

CONCLUSIONS:

Circulating tumor cells can be found in most patients with PDAC of any stage, whether localized, locally advanced, or metastatic. The ability to capture, cytologically identify, and genetically analyze CTCs suggests a possible tool for the diagnosis and characterization of genetic alterations of PDAC.

 

March, 6th
BREAKTHROUGH IN DIAGNOSIS OF PROSTATE CANCER

Dr. Sabine Mai of the Manitoba Institute of Cell Biology, CancerCare Manitoba and University of Manitoba, in collaboration with Drs. Drachenberg and Saranchuk at the Manitoba Prostate Centre, report on the use of Screencell technology for their researches on Prostate cancer.

BREAKTHROUGH IN DIAGNOSIS OF PROSTATE CANCER

WINNIPEG, MB:  Dr. Sabine Mai of the Manitoba Institute of Cell Biology, CancerCare Manitoba and University of Manitoba, in collaboration with Drs. Drachenberg and Saranchuk at the Manitoba Prostate Centre, have made advances in examining circulating tumor cells (CTCs) from the blood of prostate cancer patients. The encouraging new advancements were made possible, in part, with funds raised by the Manitoba Motorcycle Ride for Dad.

Dr. Mai, Director of the Genomic Centre for Cancer Research and Diagnosis and head of the test project, presented findings about the new blood test at a recent meeting in Boston. She co-founded 3D Signatures Inc., for commercialization of the test. “The new blood test for prostate cancer will be less invasive with a potential to be more accurate,” said Dr. Mai, who is working to get certification for the test from Health Canada.

As reported by Frank Luba in the Vancouver Province (02/03/2015), Dr. Oliver Prange (Vancouver) said the new test is focused on the intermediate group of men diagnosed with prostate cancer, which comprises about 30 per cent of the total cases. Men in the intermediate group will either receive active surveillance or aggressive cancer therapy, depending on the diagnosis. “The team looks at the structural arrangement of the chromosome component which eliminates what is a bit of a guessing game,” said Dr. Prange. “Early evidence clearly clustered patients into risk groups.  We hope to accurately predict which patients will stay indolent and which will progress,” he said.

“We hope that it will dramatically change the prognostic outlook of intermediate-risk prostate cancer patients,” said Dr. Mai.

Dr. Stuart Edmonds of Prostate Cancer Canada is encouraged by the new test. “I think it’s very promising,” Edmonds said from Toronto, where he is Prostate Cancer Canada’s vice-president of research, health promotions and survivorship. “We need to have a better test to distinguish between the aggressive disease that a man will die of rather than the more indolent, or non-aggressive disease, that a man will die with,” said Edmonds. Research is also under way to see if the new test could be used in the prognosis of other cancers, such as breast cancer, Hodgkin’s lymphoma and multiple myeloma.

Prostate cancer is the most commonly diagnosed cancer among men. It is estimated 23,600 men in Canada will be diagnosed with the cancer in 2015 and 4,000 would die from the disease.

Since 2009, over $860,000 has been raised by the Manitoba Motorcycle Ride for Dad for prostate cancer research and education. Dr. Mai’s research is proof-positive Manitobans are making a real difference. “Thank you to all Manitoba Motorcycle Ride for Dad volunteers, riders, donors and sponsors for your strong support of the project,” added Dr. Mai.

A video re: Dr. Mai’s project is found here: Dr. Sabine Mai Research Project   Visit: www.ridefordad.ca/manitoba for more information about the Motorcycle Ride for Dad.

Contacts:

Dr. Sabine Mai:                                                  (204) 787-2135  sabine.mai@umanitoba.ca

Ed Johner, Spokesperson, MRFD:                  (204) 794-5602  edjohner@icloud.com

Moe Sabourin, Co-Chair, MRFD:                    (204) 228-4301  MSabourin@wpa.mb.ca

Kirk Van Alstyne, Co-Chair, MRFD:                (204) 470-9913  kvanalstyne@winnipeg.ca

 

2014

September, 15th
An assessment of diagnostic performance of a filter-based antibody-independent peripheral blood circulating tumour cell capture paired with cytomorphologic criteria for the diagnosis of cancer.

An assessment of diagnostic performance of a filter-based antibody-independent peripheral blood circulating tumour cell capture paired with cytomorphologic criteria for the diagnosis of cancer.

Abstract

OBJECTIVES:An assessment of diagnostic performance of a filter-based antibody-independent peripheral blood circulating tumour cell capture paired with cytomorphologic criteria for the diagnosis of cancer

Circulating tumour cells (CTCs) are reported to be predictive for prognosis and response to treatment in advanced lung cancer. However, the clinical utility of the CTCs detection remains unknown for early stage lung cancer as the number of CTCs is reported as low, providing challenges in identification. We have evaluated diagnostic performance of filtration-based technology using cytomorphologic criteria in patients undergoing surgery for lung cancer.

MATERIAL AND METHODS:

We processed blood from 76 patients undergoing surgery for known or suspected lung cancer using ScreenCell(®) Cyto filter devices. Captured cells were stained using haematoxylin and eosin and independently assessed by two pathologists for the presence of atypical cells suspicious for cancer. Diagnostic performance was evaluated against pathologist reported diagnoses of cancer from surgically obtained specimens.

RESULTS:

Cancer was diagnosed in 57 patients (77.0%), including 32 with primary lung cancer (56.1%). The proportion of patients with early stage primary lung cancer in which CTCs were identified was 18 and 21 (56.3% and 65.6%, respectively) as reported by two pathologists. The agreement between the pathologists was 77.0% corresponding to a kappa-statistic of 53.7% indicating moderate agreement. No significant differences were found for the percentage of CTCs for primary and metastatic cancer as well as for cancer stages. On sensitivity weighted analysis, a sensitivity and specificity were 71.9% (95% CI 60.5-83.0) and 52.9% (95% CI 31.1-77.0), respectively. On specificity weighted analysis, a sensitivity and specificity were 50.9% (95% CI 39.3-64.4) and 82.4% (60.4-96.2), respectively.

The performance of the tested filter-based antibody-independent technology to capture CTCs using standard cytomorphologic criteria provides the potential of a diagnostic blood test for lung cancer.

An assessment of diagnostic performance of a filter-based antibody-independent peripheral blood circulating tumour cell capture paired with cytomorphologic criteria for the diagnosis of cancer

June, 13th
Colorectal carcinomas in 2014: The search for powerful prognostic markers is still on the go!

Coget J, Borrini F, Susman S, Sabourin JC. Colorectal carcinomas in 2013: The search for powerful prognostic markers is still on the go! Cancer Biomark. 2014 Jan 1;14(2):145-50.

Abstract:

Colorectal cancer (CRC) is the third cause of cancer worldwide after prostate cancer and breast cancer. Patients have a survival rate of 5 years, which varies between 10 and 95% depending on the CRC stage.

Today, the management of patients with CRC is based on parameters such as TNM and classic histologic parameters, but new molecular and cell markers have been created to improve treatment and survival. Determining the expression of a characteristic set of genes either from formalin-fixed paraffin-embedded tissues (Onco type DX test™) or from fresh tissues (AGENDIA© ColoPrint®) has led to encouraging results, but there is a need for clinical validation on a large number of patients.

Also, next-generation sequencing (NGS) technologies may be the next step in the molecular approach of CRC tumor samples, allowing tumor characterization by gene signature arrays. In addition to molecular markers, evaluation of the presence of cellular markers such as circulating tumor cells (CTC) in the blood of patients with CRC can optimize prognostic evaluation and response to treatment. CTC isolation methods used today have different sensitivities and specificities, due not only to the very small number of these cells but also to the epithelial-mesenchymal transitional process (EMT). This paper presents the preliminary results of our study conducted on CTC isolation in patients with CRC by filtration method (Screencell Cyto). This fast and efficient method identifies CTCs and also isolates cells in EMT, which explains its high efficiency compared to technologies based on immunomagnetic and microfluidic separation reliant on EpCAM presence on the cell surface.

Cancer Biomarkers 2014 Colorectal carcinomas in 2013

April, 28th
Single Cell Analysis of Circulating Tumor Cells Identifies Cumulative Expression Patterns of EMT-Related Genes in Metastatic Prostate Cancer

Chen CL, Mahalingam D, Osmulski P, Jadhav RR, Wang CM, Leach RJ, Chang TC, Weitman SD, Kumar AP, Sun L, Gaczynska ME, Thompson IM, Huang TH.Single-cell analysis of circulating tumor cells identifies cumulative expression patterns of EMT-related genes in metastatic prostate cancer. Prostate. 2013 Jun; 73(8):813-26.

Abstract

BACKGROUND: Prostate tumors shed circulating tumor cells (CTCs) into the blood stream. Increased evidence shows that CTCs are often present in metastatic prostate cancer and can be alternative sources for disease profiling and prognostication. Here we postulate that CTCs expressing genes related to epithelial-mesenchymal transition (EMT) are strong predictors of metastatic prostate cancer.

METHODS: A microfiltration system was used to trap CTCs from peripheral blood based on size selection of large epithelial-like cells without CD45 leukocyte marker. These cells individually retrieved with a micromanipulator device were assessed for cell membrane physical properties using atomic force microscopy. Additionally, 38 CTCs from eight prostate cancer patients were used to determine expression profiles of 84 EMT-related and reference genes using a microfluidics-based PCR system.

RESULTS: Increased cell elasticity and membrane smoothness were found in CTCs compared to noncancerous cells, highlighting their potential invasiveness and mobility in the peripheral circulation. Despite heterogeneous expression patterns of individual CTCs, genes that promote mesenchymal transitioning into a more malignant state, including IGF1, IGF2, EGFR, FOXP3, and TGFB3, were commonly observed in these cells. An additional subset of EMT-related genes (e.g., PTPRN2, ALDH1, ESR2, and WNT5A) were expressed in CTCs of castration-resistant cancer, but less frequently in castration-sensitive cancer.

CONCLUSIONS: The study suggests that an incremental expression of EMT-related genes in CTCs is associated with metastatic castration-resistant cancer. Although CTCs represent a group of highly heterogeneous cells, their unique EMT-related gene signatures provide a new opportunity for personalized treatments with targeted inhibitors in advanced prostate cancer patients.

Copyright © 2012 Wiley Periodicals, Inc.

Chen, et. al The Prostate

April, 28th
Rapid Separation of Mononuclear Hodgkin from Multinuclear Reed-Sternberg Cells

Lab Hematol 2014 Rapid Separation of Mononuclear Hodgkin from Multinuclear Reed-Sternberg Cells

Kongruttanachok N, Cayre YE, Knecht H, Mai S. Rapid separation of mononuclear hodgkin from multinuclear reed-sternberg cells. Lab Hematol. 2014 Mar 1;20(1):2-6.

We describe a method to isolate small mononucleated Hodgkin (H) cells from multinucleated Reed Sternberg (RS) cells of Hodgkin lymphoma using the ScreenCell filter device. This filtration-based approach lends itself to future clinical applications in that it enables the separation of H and RS cells from lymph node biopsies, bone marrow aspirates, pleural effusions, and blood, including the isolation of monoclonal Hodgkin precursor cells from the blood.

April, 25th
3D nuclear telomeric signatures define circulating tumor cells (CTCs) and characterize CTC subpopulations in intermediate risk prostate cancer patients

Awe, et.al. AACR April2014_pdf

2013

September, 19th
Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer ?

El-Heliebi A, Kroneis T, Zöhrer E, Haybaeck J, Fischereder K, Kampel-Kettner K, Zigeuner R, Pock H, Riedl R, Stauber R, Geigl JB, Huppertz B, Sedlmayr P, Lackner C. Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer? J Transl Med. 2013 Sep 17; 11(1):214.

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Abstract

BACKGROUND: Single circulating tumor cells (CTCs) or circulating tumor microemboli (CTMs) are potential biomarkers of renal cell cancer (RCC), however studies of CTCs/CTMs in RCC are limited. In this pilot study we aimed to evaluate a novel blood filtration technique suited for cytomorphological classification, immunocytochemical and molecular characterization of filtered, so called circulating non-hematologic cells (CNHCs) – putative CTCs/CTMs – in patients with RCC.

METHODS: Blood of 40 patients with renal tumors was subjected to ScreenCell filtration. CNHCs were classified according to cytomorphological criteria. Immunocytochemical analysis was performed with antibodies against CD45, CD31 and carbonic anhydrase IX (CAIX, a RCC marker). DNA of selected CNHCs and respective primary tumors was analysed by array-CGH.

RESULTS: CNHC-clusters with malignant or uncertain malignant cytomorphological features – putative CTMs – were negative for CD45, positive for CD31, while only 6% were CAIX positive. Array-CGH revealed that 83% of malignant and uncertain malignant cells did represent with a balanced genome whereas 17% presented genomic DNA imbalances which did not match the aberrations of the primary tumors. Putative single CTCs were negative for CD45, 33% were positive for CD31 and 56% were positive for CAIX.

CONCLUSIONS: The majority of CNHC-clusters, putative CTMs, retrieved by ScreenCell filtration may be of endothelial origin. Morphological criteria seem to be insufficient to distinguish malignant from non-malignant cells in renal cancer.

September, 19th
Detection of circulating tumor cells in patients with adrenocortical carcinoma: a monocentric preliminary study

Pinzani P, Scatena C, Salvianti F, Corsini E, Canu L, Poli G, Paglierani M, Piccini V, Pazzagli M, Nesi G, Mannelli M, Luconi M. Detection of circulating tumor cells in patients with adrenocortical carcinoma: a monocentric preliminary study. J Clin Endocrinol Metab. 2013 Sep; 98(9):3731-8.

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Abstract:

CONTEXT: Adrenocortical carcinoma (ACC) is a rare malignancy, the prognosis of which is mainly dependent on stage at diagnosis. The identification of disease-associated markers for early diagnosis and drug monitoring is mandatory. Circulating tumor cells (CTCs) are released into the bloodstream from primary tumor/metastasis. CTC detection in blood samples may have enormous potential for assisting in the diagnosis of malignancy, estimating prognosis, and monitoring the disease.

OBJECTIVE: The aim of the study was to investigate the presence of CTCs in blood samples of patients with ACC or benign adrenocortical adenoma (ACA).

INTERVENTION: CTC analysis was performed in blood samples from 14 ACC patients and 10 ACA patients. CTCs were isolated on the basis of cell size by filtration through ScreenCell devices, followed by identification according to validated morphometric criteria and immunocytochemistry. We measured the difference in CTC detection between ACC and ACA.

RESULTS: CTCs were detected in all ACC samples, but not in ACA samples. Immunocytochemistry confirmed the adrenocortical origin. When ACC patients were stratified according to the median value of tumor diameter and metastatic condition, a statistically significant difference was found in the number of CTCs detected after surgery. A significant correlation between the number of CTCs in postsurgical samples and clinical parameters was found for tumor diameter alone.

CONCLUSIONS: Our findings provide the first evidence for adrenocortical tumors that CTCs may represent a useful marker to support differential diagnosis between ACC and ACA. The correlation with some clinical parameters suggests a possible relevance of CTC analysis for prognosis and noninvasive monitoring of disease progression and drug response.

February, 19th
Three-Dimensional Telomeric Analysis of Isolated Circulating Tumor Cells (CTCs) Defines CTC Subpopulations.

Adebayo Awe J, Xu MC, Wechsler J, Benali-Furet N, Cayre YE, Saranchuk J, Drachenberg D, Mai S. Three-Dimensional Telomeric Analysis of Isolated Circulating Tumor Cells (CTCs) Defines CTC Subpopulations. Transl Oncol. 2013 Feb; 6(1):51-65.

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January, 19th
Usefulness of circulating tumor cell detection in pancreatic adenocarcinoma diagnosis

Iwanicki-Caron I, Basile P, Toure E, Antonietti M, Lecleire S, Di Fiore A, Oden-Gangloff A, Blanchard F, Lemoine F, Di Fiore F, Sabourin JC, Michel P. Usefulness of circulating tumor cell detection in pancreatic adenocarcinoma diagnosis. Am J Gastroenterol. 2013 Jan;108 (1):152-5.

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2011

February, 19th
A new device for rapid isolation by size and characterization of rare circulating tumor cells.

Desitter I, Guerrouahen BS, Benali-Furet N, Wechsler J, Jänne PA, Kuang Y, Yanagita M, Wang L, Berkowitz JA, Distel RJ, Cayre YE. A new device for rapid isolation by size and characterization of rare circulating tumor cells. Anticancer Res. 2011 Feb; 31(2):427-41.

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