Methods for diagnosis of myelodysplastic syndromes (MDS)

• US 8,377,649 B2
• Filed: 02/01/2011
• Issued: 02/19/2013
• Est. Priority Date: 11/09/2004
• Status: Active Grant

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Moore, Jonni, Cherian, Sindhu, Bagg, Adam
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US13/019,167
• Publication Number:
  US 20120028276A1
• Time in Patent Office: 749 Days
• Field of Search: 435/2, 435/7.2, 435/7.24, 435/7.25, 435/287.2, 435/973, 436/518, 436/523, 436/525, 436/526, 436/56, 436/63, 436/64, 422/50, 422/61, 422/73, 422/82.08, 422/82.09, 422/101
• US Class Current: 435/7.23
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: G01N 2333/70553 Integrin beta2-subunit-containing molecules, e.g. CD11, CD18
• CPC Class Codes: G01N 2333/70589 CD45
• CPC Class Codes: G01N 2800/22 Haematology
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80
• CPC Class Codes: Y10S 435/973 Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10S 436/813 Cancer
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]
• CPC Class Codes: Y10T 436/13 Tracers or tags

Abstract

The present invention relates to methods and kits for diagnosing, ascertaining the clinical course of myelodysplastic syndrome (MDS) and ascertaining response to a therapy regimen of myelodysplastic syndrome. Specifically the invention provides methods and kits useful in the diagnosis and determination of clinical parameters associated with MDS based on surface markers unique to MDS.

Claims

1. A method of ascertaining response to a therapy regimen in a subject diagnosed with myelodysplastic syndrome, comprising:

obtaining a blood sample from the subject at a timepoint during or after said therapy regimen;

measuring and analyzing a set of predictive parameters in leukocytes from the blood sample of the subject using flow cytometry, wherein said set of predictive parameters comprises polymorphic neutrophil (PMN) granularity and leucocyte expression of CD66, CD11a, CD10, and CD116;

assigning a numerical value to each predictive parameter;

assigning a numerical score reflecting the predictive parameter values obtained for said leucocytes in said blood sample;

comparing the predictive parameters to corresponding predictive parameters for a control sample taken from said subject obtained prior to the initiation of, or earlier than said timepoint in, said therapy regimen, or a combination thereof;

and assigning a numerical score reflecting predictive parameter values obtained for the control sample, wherein a decrease in the subject'"'"'s numerical score in comparison to the control sample indicates the subject'"'"'s responsiveness to said therapy regimen.

2. The method of claim 1, wherein said score reflects the variance between the mean of values obtained for said predictive parameters of said leucocytes in said blood sample from that of said control sample.

3. The method of claim 1, wherein said predictive parameters further comprise side scatter capability.

4. The method of claim 3, wherein said neutrophil granularity is measured by evaluating CD45 expression as a function of side scatter of said PMN.

5. The method of claim 1, further comprising the step of lysing red blood cells from said blood sample.

6. The method of claim 1, wherein said blood sample is enriched for leukocytes.

7. The method of claim 1, wherein said blood sample is enriched for neutrophils in said sample.

8. The method of claim 1, wherein said myelodysplastic syndrome (MDS) is refractory cytopenia with multilineage dysplasia (RCMD) refractory anaemia (RA), acquired idiopathic sideroblastic anaemia (AISA), refractory anaemia with ring sideroblasts (RARS), refractory anaemia with excess blasts (RAEB), refractory anaemia with excess blasts in transformation (RAEB-t), chronic myelomonocytic leukaemia (CMML), atypical chronic myelogenous leukemia (aCML), 5q-syndrome diseases, which are erythroid dysplasia, thrombocytosis, or hypolobated micromegakaryocytic hyperplasia, or a combination thereof.