Download karen anderson AZMN 2014

When B Cells Go Bad: Infection,
Inflammation and Chronic B
Cell Stimulation
Karen S. Anderson MD PhD
Associate Professor, Biodesign Institute
Arizona State University
Mayo Clinic Arizona
Conflicts of Interest
• I serve on the advisory board and have received consultant
fees and stock options with Provista Dx.
• No off-label clinical diagnostics or therapeutics will be
discussed.
W. Michael Kuehl & P. Leif Bergsagel, 2002
Monoclonal Gammopathy of Undetermined Signficance
• 3.2% of adults over the age of 50; 6.6% of adults over age 80
• Premalignant disease that can transition to myeloma; average
2-15 years, rate 1% per year
• Over 90% of patients with MM have a premalignant plasma
cell disorder
• 50% Ig translocations; 50% hyperdiploid
– Errors in switch recombination and somatic hypermutation of the B
cell
• Likely due to an abnormal response to antigenic stimulation
Chronic Infections and Antibodies
• EBV, HHV-8, HCV, (CMV), H. pylori are associated with B cell
lymphoma and B cell chronic lymphocytic leukemia
– what about myeloma?
• Ig in HCV-positive myeloma patients can target the virus
• ~20% of MGUS and myeloma Abs target infectious antigens
(ASH 2013)
• Would treatment of chronic infections also prevent or treat
MGUS?
Chronic Infections not Associated with MGUS
Bida and Rajkumar Mayo Clin Proc 2009
How B cells Proliferate in response to infection
Moir and Fauci, Nat Rev Immunol 2009
Antibody structure:
Ligands (antigens)
• What is the specificity of antibodies?
• What is the diversity of antibodies?
When Ig genes rearrange, there are >1011 potential molecules
To facilitate research, DNASU stores over 162,000 plasmids and distributes these to researchers
in 37 states and 38 countries
Nucleic Acid-Programmable Protein Array (NAPPA)
Proteomics: the study of all of the proteins in the human body
We use plasmids to make over 10,000 human proteins
By putting these plasmids on a glass slide (NAPPA) we can make and study over 10,000 proteins
at one time
1. Print Plasmids
Protein
Expression
…
•
•
•
Replicate arrays of candidate proteins
2. Express & capture
proteins
Find antibodies in patients’ blood
NAPPA Array Production and Screening
Gene Cloning
Bacterial plating
DNA preparation
Array Printing
Antigen array
Add patient
serum
p53
Serologic Biomarkers for HPV+ Cancer
Human papilloma virus (HPV16): ~70% of oropharyngeal cancers
Emerging epidemic in US and Europe
HPV Genome: 8 ORFs
Diagnosis
Prognosis
NE2, p<0.001
0.75
0.50
0.25
NE2 positive
NE2 negative
0.00
Proportion surviving
1.00
Kaplan-Meier survival estimates
0
12
Number at risk
NE2 positive 84
NE2 negative 13
75
9
24
36
48
Follow-up time (months)
66
6
59
4
33
3
60
10
1
Identification of the Targets of Antibodies
• Protein microarrays are now used to test >10,000 proteins for
antibody targets
• Large gene collections can be leveraged for rapid protein display
• We are developing pathogen-specific arrays to identify
antibodies in blood
• We need to measure the antibody IMMUNOME to understand the
pathogenesis of MGUS
What is the diversity of Antibodies?
Can we detect specific Ig rearrangements?
DNA Origami nanostructure design
Hao Yan and Joe Blattman, Biodesign
Overall strategy to obtain linked RNA sequences from single cells.
Conclusions
 Antibodies have extraordinary sequence diversity
 There are emerging technologies for quantitating that diversity
 By linking single-cell RNA capture with next-gen sequencing, we
may be able to:
 Identify early events (?pre-MGUS) of loss of diversity
 Rapidly generate patient-specific probes for molecular detection of
rearrangements
Acknowledgements
ASU Biodesign Institute
Anderson Lab
 Joseph Blattman
 Julia Cheng
 Hao Yan
 Ting Li
 Josh LaBaer
 Rizwan Alam
 Benjamin Katchman  Ji Qiu
 Krishna Sundaresan  Garrick Wallstrom
 Laura Gonzalez
 Diego Chowell
 Jin Park
 Shay Ferdosi
 Fernanda Festa
 Hans Frykman
 I.Purushothaman
 Fernando Hernandez
Mayo Oncology
 Robert Brown
 Don Northfelt
 Alison Goulder
 Doug Lake
 Jack Resnik
 Barb Pockaj
 Peter Chang
 Michael Barrett
Our Patients
NCI/Early Detection Research Network
Zicarelli Foundation