NCI Clinical Flow Cytometry Unit

Constance M. Yuan, M.D. Ph.D

Primary Research Area(s)

Expertise: AP/CP Board Certified, Hematology Board Certified.
Focus of expertise in Multiparametric Clinical Flow Cytometry (FCM) Instructor for the Flow Cytometry Course at the International Clinical Cytometry Society (ICCS), which includes participants from UK, Western Europe, India, East Asia, Australia, South America, as well as Canada and the U.S. Minimal Residual Disease Detection in Myeloma Consensus Initiative Member
Serving on Panel of Experts on Aug 21, held here at NIH.

Flow Cytometry Consultant/Liason to DLM Hematology, Volunteer for Bone marrow service for DLM Hematology.
Service work has included coverage of Hematopathology related areas (sign-out of bone marrow biopsies, bone marrow aspirate, body fluids, lymph nodes) as well as Coagulation and Molecular-related areas (B and T-cell clonality).

What technologies / approaches / methodologies do you utilize?

Multiparametric Flow Cytometry 8 color clinical flow immunophenotyping Detection of CAR19 T-cells, and T-cell subset detection Antigen Quantitation in the form of Antibody Binding Capacity TCR Vbeta Repertoire analysis for T-cell clonality studies MRD Applied to ALL, CLL, HCL, AML, and now PCM MRD approach is applicable to MGUS, the precursor state of PCM.

Data/Examples
New Markers for Clinical detection of disease PCM Use of CD81 improves detection of PCM MRD (95% sensitivity, 100% specificity).
Improvements in performing flow cytometry for PCM in BM Immunophenotyping of bilateral BM study in PCM (Manasanch E et al, Abstract, ASCO 2013).
13.79%
10
5

SF13 931 pb MGUS rpt_05_B-8.fcs com pensated FSC SSC PCs via

0.00%

CD56 PC7

10

10

10

10.34%
10
2

75.86%
10 10 CD19 APC
3 4

10

Mailankody S, et al. Minimal residual disease (MRD) detection by deep sequencing in newly diagnosed multiple myeloma patients treated with carfilzomib, lenalidomide and dexamethasone BM or plasma samples from 14 MM patients treated with CRd, subjected to deep sequencing to detect a high frequency myeloma-specific gene rearrangement. MRD subsequently assessed by sequencing and by marrow flow cytometry. Result: 7/13 patients achieved VGPR/CR/nCR based on traditional protein response criteria. 1/7 was (+) by sequencing and (-) by FCM. 1/7 was (+) by FCM and (-) by sequencing. FCM may potentially be within the realm of detection sensitivity of MRD in PCM as sequencing!

Data/Examples
Table 1. Flow cytometry of the bone marrow to detect MRD in multiple myeloma: Current practice based on a survey of 30 major institutions in the U.S.

Blood, Aug 8 2013, Vol 122, No. 6.

Research Implications
How does your research fit in the broader context? How does it fit into LP and broader areas? Within LP: Integrates with Cytology (CSF, FNA, other body fluids), Cytogenetics (BM aspirate), Hematopathology, and Molecular (morphologic and clonality correlation). Broader areas include DLM: Further morphologic correlation, working conferences directly with different clinical groups (myeloma group, transplant group).
How does your research translates into potential clinical applications?
Physicians use our data for diagnosis, but also to guide choice of therapy often in real time, for protocol enrollment, and to decide between protocols or if protocol switch is needed. Our data provides an assessment of efficacy (or lack of efficacy) of new therapeutics. Regarding autologous stem cell collections, we detect whether there is disease contaminating the products.

Research Implications
PCM MRD by flow is rapid, and detects disease earlier and more sensitively than traditional methods (M-spike, molecular, bone marrow biopsy PC count, CD138 IHC). MRD detection, prognosis post transplant, plus early treatment with new PCM drugs. Potential for curability of a previous incurable disease. FCM has potentially has the same level of sensitivity as deep sequencing. Potential for routine clinical applications.

Future Direction
Where do you see your research going in the next 5-10 years?
Continue to developing new testing to assess efficacy of novel therapeutics Working directly with collaborators, currently in planning stage.

What are the current challenges & obstacles?

Having to work in an environment (physical, administrative, budgetary, etc.) that may not be optimally designed for the nature of the work that we do.

But we do our best And try to remain flexible And strive to be creative With what we have!

Collaborators
LP, NCI Cytogenetics Diane Arthur Cytology Mark Roth Arma ndo Filie Hematopathology Stefania Pittaluga Elaine Jaffe Molecular Mark Raffeld Myeloma Group MOB, NCI: Ola Landgren, Mark Roschewski CLL Group NHLBI: Adrian Wiestner Andy Rawstron, Leeds Teaching Hospital, UK

Pediatric Group POB, NCI: Crystal Mackall, Terry Fry, Trey Lee Alan Wayne, Childrens Hospital, L.A.
Transplant ETIB, NCI: Claude Sportes Hairy Cell Leukemia Group LMB, NCI: Robert Kreitman, Ira Pastan

Hematology DLM, CC Kathy Calvo Roger Kurlander Irina Maric Raul Braylan