biocept scientific publications

Biocept Publications, Studies, Posters and Abstracts.

Conclusions:

  1. Study demonstrates that the peripheral blood of cancer patients contain circulating tumor cells other than those normally detected with antibodies to EpCAM and cytokeratin.
  2. The use of CEE-Enhanced and antibody mixtures along with the traditional anti-EpCAM and anti-CK approach may lead to new insight into the diagnostic applications of CTCs.

Stephen D.Mikolajczyk1, Lisa S.Millar1, Pavel Tsinberg1, Stephen M. Coutts1, Maryam Zomorrodi1, Tam Pham2, Farideh Z. Bischoff2, and Tony J. Pircher1

1Research and Development, Biocept Inc., 5810 Nancy Ridge Drive, Suite 150, San Diego, CA 92121, USA
2Translational and Clinical Development, Biocept Inc., 5810 Nancy Ridge Drive, Suite 150, San Diego, CA 92121, USA

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Conclusions:

  1. The cell enrichment and extraction microfluidic platform (OncoCEETM) provides a sensitive approach for evaluation of HER2 in CTCs and DTCs.
  2. CTCs and DTCs acquired HER2 gene amplification in 21% and 7% of patients with HER2 negative early stage primary breast cancer.
  3. CTCs and DTCs lost HER2 gene amplified status in 57% and 83% of patients with HER2 positive early stage primary breast cancer.
  4. The clinical significance of alterations in HER2 status among CTCs and DTCs in early stage breast cancer needs further investigation.

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Conclusions:

  1. The CEETM technology provides a sensitive platform for enhanced capture, detection and molecular characterization (ER/PR) in intact CTCs within the microchannels .
  2. Sensitivity and accuracy levels of this test need to be tested on a larger patient population.
  3. Though some discordance between tumor and CTCs is expected given variation in tumor heterogeneity, biopsy size, and robustness of the technical assay (especially for IHC), a blood-based CTC assay may offer more reliable testing given the advantages of simple repeat testing and the use of larger blood volumes, which may help to ensure informative results.

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Conclusions:

  1. It is feasible to use deep sequencing to detect rare somatic mutations in highly heterogeneous enriched CTC samples.
  2. We are comparing the mutational profile between CTCs and primary/metastatic tumors (ongoing).
  3. Novel mutations associated with tumorigenesis and/or metastasis may be explored using this method.

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Conclusions:

  1. Using a HER2/CEP17 cut-off of 2.0, HER2 gene amplified CTCs and/or DTCs were detected in 4/15 (27%) patients classified as HER2+ in primary tumor.
  2. Among HER2- primary tumor patients (n=92), HER2 amplified CTCs were detected in 6 (6.5%) and DTCs in 18 (20%) cases.
  3. An overall HER2 discordance rate of 25% (27 of 107 cases) was observed (based on HER2:CEP17 ratio of 2.0). The discordance rate is lowered to 18% when a HER2:CEP17 ratio of 2.2 is applied.
  4. Discordant HER2 status was contributed mainly by HER2+ DTCs occurring in patients with HER2- primary breast tumors.
  5. The clinical significance of evaluating the status of HER2 gene amplification in CTCs and DTCs in the management of patients with breast cancer needs to be evaluated prospectively in larger clinical trials to assess efficacy in treating patients classified as HER2+ by CTC/DTC analysis

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Conclusions:

There is significant heterogeneity between ER/PR protein expression in CTCs and primary tumor/metastatic biopsy and this status may change over time due to therapy. ER/PR ICC on CTCs from peripheral blood using the OncoCEETM platform is shown to be feasible with high concordance (86%) in ER/PR status between primary tumor/metastatic biopsy (by IHC) and CTCs (by ICC). The significance of heterogeneity at the ER/PR protein level in CTCs ascertaining to the prognosis and predictive response to anti-estrogen therapy needs further evaluation in larger prospective clinical trials.

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Conclusions:

  1. The cell enrichment and extraction (CEE™) microfluidic technology provides a sensitive platform for evaluation of HER2 by FISH in intact CTCs and DTCs.
  2. HER2+ CTCs and DTCs were encountered in patients with HER2+ as well as HER2- primary tumors.
  3. Discordant HER2 status was contributed mainly by HER2+ DTCs occurring in HER2- primary breast tumors.
  4. The clinical implications of evaluating HER2 in CTCs and DTCs in patients with invasive breast cancer needs further investigation.

S Krishnamurthy1, FZ Bischoff2, K Wong2,S Mikolajczyk2 , T Pham2 , H.M. Kuerer, A Lodhi3 , A Bhattacaryya, C Hall3 , and A Lucci3
Universityof TexasMD Anderson Cancer Center Department ofPathology 1, SurgicalOncology3, Biocept, Inc., San Diego CA2

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Conclusions:

  1. CEE™ technology provides a sensitive platform for enhanced capture, detection, and characterization of both CK+ and CK- CTCs.
  2. This platform allows for evaluation of HER2 gene amplification status by FISH in intact CTCs within the microchannels at sensitivity and accuracy levels suitable for standardized CLIA-laboratory testing.

Julie Ann Mayer, PhD, Tony J. Pircher, PhD, Stephen D. Mikolajczyk, Philip D Cotter, PhD, FACMG, Farideh Z. Bischoff, PhD,Biocept Inc, San Diego, CA 92121

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Conclusions:

  1. We have developed a robust method of detecting CTCs that can capture both epithelial and mesenchymal phenotypes.
  2. Our findings suggest current enrichment techniques may be missing an important population of CTCs.

Chad V. Pecot1, Farideh Bischoff2, Yvonne Lin3, Padmavathi Jaladurgam1, William Merritt4, Tony Pircher2, Steve Mikolajczyk2, Julie Mayer2, Karina Wong2, Tam Pham2, Justin Bottsford-Miller1, Rebecca Stone1, Joseph Celestino1, Alpa Nick1, Cathy Eng1, Anil Sood1

1UT M.D. Anderson Cancer Ctr., Houston, TX
2Biocept Incorporated, San Diego, CA;
3University of Southern California, Los Angeles, CA
4South Carolina Oncology, Columbia, SC

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Conclusions:

  1. CEE™ technology provides a sensitive platform for enhanced capture, detection, and characterization of both CK+ and CK- CTCs.
  2. This platform allows for evaluation of HER2 gene amplification status by FISH in intact CTCs within the microchannels at sensitivity and accuracy levels suitable for standardized CLIA-laboratory testing.

Julie Ann Mayer, PhD, Tony J. Pircher, PhD, Stephen D. Mikolajczyk, Philip D Cotter, PhD, FACMG, Farideh Z. Bischoff, PhD,
Biocept Inc, San Diego, CA 92121

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Conclusions:

The CEE-Enhanced staining technology enables the detection of CK-negative CTCs. This novel detection method, based on the in situ labeling of antibodies used for capture, greatly expands single and multi-antibody approaches to the study of rare circulating cells. Specifically, this allows the exploration and study of circulating cells not expressing CK such as highly de-differentiated tumor cells, stem cells or those tumor cells undergoing EMT. The clinical significance of these CK-negative CTCs is under investigation.

Stephen D. Mikolajczyk, Lisa S. Millar, Maryam Zomorrodi, Farideh Z. Bischoff, Jayne Scoggin, Tam Pham, Karina Wong, Tony J. Pircher. Biocept, Inc., San Diego, CA

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Conclusions:

  1. There is a significant heterogeneity between the gene amplification status and protein overexpression of ESR1.
  2. The gene status of ESR1 ranges from negative, equivocal and amplified in both ER negative and ER immunopositive cases.
  3. The significance of heterogeneity at the ESR1 gene locus in ascertaining the prognosis and predictive response to antiestrogen therapy needs further evaluation in larger prospective clinical trials.

Julie A. Mayer1, Tam Pham1, Karina Wong1, Anthony Lucci2, Farideh Bischoff1, Savitri Krishnamurthy2.
1Biocept Inc, San Diego, CA;
2The University of Texas MD Anderson Cancer Center, Houston, TX

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Conclusions:

  1. Antibody mixtures improve the recovery of cancer cells, including CTCs not captured with anti-EpCAM alone.
  2. CEE-Enhanced™ can be used to stain additional CTCs that do not stain with anti-cytokeratin.
  3. CEE™ technology allows multiple screening and staining strategies for the analysis of CTCs.

Stephen D. Mikolajczyk, Lisa S. Millar, Pavel Tsinberg, Maryam Zomorrodi, Jayne Scoggin, Tam Pham, Karina Wong, Farideh Z. Bischoff, Tony J. Pircher
Biocept Inc, San Diego, CA 92121

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Conclusions:

  1. The cell enrichment and extraction microfluidic technology (CEE) provides a sensitive platform for enhanced detection and characterization of antibody cytokeratin positive and negative CTCs.
  2. This platform allows evaluation of HER2 gene amplification status by fluorescence in situ hybridization (FISH) in intact CTCs within the microchannels.
  3. The utility of this platform for phenotypic and genotypic characterization of CTCs in breast cancer needs to be tested in larger clinical trials.

Savitri Krishnamurthy1, Farideh Z. Bischoff2, Steve Mikolajczyk2,and Anthony Lucci3
1University of Texas MD Anderson Cancer Center Department of Pathology
2Biocept, Inc., San Diego CA
3Surgical Oncology

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Conclusions:

  1. Extremely rare cells can be separated with high efficiency and high purity for in situ inspection.
  2. Higher antigen densities yield higher capture. Middle flow rates yield constant k values, avoiding streamline effect.

Mary Dickson1,2, Pavel Tsinberg1, Zhongliang Tang1, Timothy Wilson1, Edward Leonard2
1Biocept, Inc., San Diego, CA
2Department of Chemical Engineering, Columbia University, New York, NY

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Conclusions:

Standard marker and FISH analysis of the cells can be completed within the device for highly accurate diagnosis.

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Conclusions:

Farideh Z. Bischoff1, Pavel Tsinberg1, William M. Merritt2, Yvonne G. Lin2, Anil K. Sood2
1Biocept, Inc., San Diego, CA
2MD Anderson Cancer Center, Dept of Gynecologic Oncology, Houston, TX, Department of Urology, Houston, TX

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Conclusions:

Michael Williams1, Tony Pircher2, Maryam Zomorrodi2, Steve Mikolajczyk2, Ashish Kamat1

1Urology, MD Anderson Cancer Center
2Biocept

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Conclusions:

  1. Overall, data concluded that the CEE system conforms to simple physical principles and that it robustly and efficiently captures tumor cells under a variety of operating conditions.
  2. There is a clear two step recommendation. First, clinicians should employ antibody cocktails with a common capture principle, here biotin, in order to maximize relevant cell capture. Second, while the data presented here show that in most cases with sufficient surface antibody density, greater than 70% capture was achieved, a channel design which forces variations in flow velocities along the channel length may overcome potential inefficiencies in capture seen at low antigen densities.

Mary Nora Dickson1,2, Pavel Tsinberg1, Zhongliang Tang1, Farideh Z. Bischoff1, Timothy Wilson1, and Edward F. Leonard2
1Biocept, Inc., 5810 Nancy Ridge Drive Suite 150, San Diego, California 92121, USA
2Columbia University School of Engineering and Applied Science, Department of Chemical Engineering

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Conclusions:

  1. Current assays for CTC capture likely miss populations of cells that have undergone EMT. Capture and study of CTCs that have undergone EMT would allow a better understanding of the mechanisms driving metastasis.
  2. The results confirm the utility of our microfluidics platform as a reliable method for assay development and efficient recovery of CTCs. We observed that a potentially important population of cancer cells is present in circulation that would likely be missed by standard detection criteria.

Chad V. Pecot, Farideh Z. Bischoff, Julie Ann Mayer, Karina L. Wong, Tam Pham, Justin Bottsford-Miller, Rebecca L. Stone, Yvonne G. Lin, Padmavathi Jaladurgam, Ju Won Roh, Blake W. Goodman, William M. Merritt, Tony J. Pircher, Stephen D. Mikolajczyk, Alpa M. Nick, Joseph Celestino, Cathy Eng, Lee M. Ellis, Michael T. Deavers, and Anil K. Sood
Cancer Discovery December 2011 1:580-586; Published OnlineFirst November 3, 2011; doi:10.1158/2159-8290.CD-11-0215

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Conclusions:

  1. OncoCEE-BR allows for evaluation of HER2 gene amplification status by FISH in intact CTCs within the microchannels at sensitivity and accuracy levels suitable for standardized CLIA-laboratory testing:
    Concordance between CTCs and the primary Tumor = 93%
    Overall test sensitivity = 95%
    Overall test specificity = 92%
  2. The CEETM technology provides a sensitive platform for enhanced capture, detection and characterization of CTCs (CK+/CD45-/DAPI+) that performs better than Veridex.
  3. The OncoCEE-BR test with HER2 could be utilized to confirm HER2 tumor tissue testing, especially in tissue-negative cases, to ensure that patients suitable for trastuzumab therapy are not inadvertently missed.

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Conclusions:

  1. Using Biocepts’ OncoCEE platform, discordance in HER2 status was observed among CTCs (15%) and DTCs (28.8%) as compared to primary tumor biopsy.
  2. Among patients with a HER2- primary tumor, 5 of 79 (6.3%) and 14 of 67 (20.8%) had HER2+ CTCs and DTCS, respectively, suggesting a significant number of patients may be potential candidates for Herceptin therapy.
  3. CTCs (blood) and DTCs (bone marrow) may serve as an alternative source of tumor tissue to aid in the determination and/or verification of HER2.
  4. HER2 gene amplification in circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) may be useful for modifying Herceptin therapy in breast cancer.

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Conclusions:

  1. Selector Assay suppresses wild-type amplification by greater than 100,000 fold
  2. Has little to no suppresive effect on the amplification of mutant alleles.
  3. Mutations are detected in a wild-type background at better than 1:10,000.
  4. The presence of a wild-type allele at greater than 2000 fold excess in a complex genomic background has no adverse effect on mutant allele amplification, detection or quanitfication
  5. Works with both DNA and RNA targets from clinical samples
  6. Demonstrated utility of the T790M Selector assay in NSCLC patient samples
  7. Works in real-time, end-point, and melt-curve analysis. Seamlessly interfaces to sequencing and other confirmatory methods of analysis once mutant alleles are selectively amplified.

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