The document reports on a study that investigated the "Dutch hypothesis" and "British hypothesis" about asthma and COPD. Biological clustering of measurements from 86 asthma and 75 COPD patients supported both hypotheses. It identified clusters of patients with distinct asthma and COPD features, as well as overlapping clusters, validating both hypotheses. The study was then validated on a new group of 166 asthma and 58 COPD patients using two different approaches.

1. Biological clustering supports both Dutch and
British hypotheses of Asthma and COPD
Michael A Ghebre
University of Leicester
III Annual PGR Conference
University of Leicester, UK
31st March 2014

2. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Table of contents
1 Introduction and Objectives
2 Asthma and COPD Biological Clustering
3 Asthma and COPD Validation Study
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
4 Summary and Conclusion
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

3. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Table of contents
1 Introduction and Objectives
2 Asthma and COPD Biological Clustering
3 Asthma and COPD Validation Study
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
4 Summary and Conclusion
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

4. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

5. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

6. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

7. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

8. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

9. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

10. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

11. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

12. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

13. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

14. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

15. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

16. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Introduction and Objectives
Severe asthma and chronic obstructive pulmonary disease (COPD)
are complex and heterogeneous diseases.
”Dutch hypothesis” => Asthma and COPD are the same disease
with the same underlying mechanism.
”British hypothesis” => Asthma and COPD are two distinct
disease with di↵erent underlying mechanism.
We investigated whether they represent distinct or overlapping
conditions in terms of their available measurements
We do have access to a large number of measurements on 86
asthma and 75 COPD subjects; i.e.
Demographic characteristics
Clinical and lung function
Biological characteristics (i.e sputum cytokines)
This study was validated on an independent severe asthma (n=166)
and COPD (n=58) using two approaches.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

17. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary statistics of some characteristics...
Table 1. Demographic, lung function and clinical characteristics across Asthma and COPD
Variable Asthma=86 COPD=75 P-value
Male [n(%)] 43 (50) 53 (70.7) 0.008
Current or Ex- smokers [n (%)] 32 (37.2) 72 (96.0) < 0.0001
Pack -year history 4.6 (2.98 - 7.26) 40 (34.46 - 46.39) < 0.0001
Age (years)+
54 (1.3) 69 (1.1) < 0.0001
Duration of Disease (years) 21 (16.4 - 26.5) 5 (4.12 - 6.55) < 0.0001
BMI (kg/m2
)+
30.4 (0.8) 25.7 (0.5) < 0.0001
Pre FEV1/FVC ratio (%)+
67.6 (1.5) 49.8 (1.6) < 0.0001
Pre FEV1 Predicted (%)+
74.6 (2.4) 45.4 (2.1) < 0.0001
Post FEV1 Predicted (%)+
79.8 (2.4) 47.1 (2.1) < 0.0001
Sputum Neutrophil count (%)+
63.2 (2.5) 69.7 (2.5) 0.07
Sputum Eosinophil count (%)+
2.1 (1.38 - 3.1) 1.4(0.98 - 1.93) 0.14
Sputum Macrophage count (%)+
16.7 (13.41 - 20.78) 16.2 (13.3 - 19.8) 0.84
Data presented as Geometric mean with corresponding confidence interval unless stated, +
Mean (SEM)
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

18. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Cytokines across Asthma and COPD
Figure : A=Higher in Asthma and B=Higher in COPD
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

19. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Where Asthma(A) and COPD (C) overlap?
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

20. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Table of contents
1 Introduction and Objectives
2 Asthma and COPD Biological Clustering
3 Asthma and COPD Validation Study
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
4 Summary and Conclusion
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

21. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
Cluster analysis is unsupervised statistical techniques which identifies
homogeneous subgroups and creates labels for further supervised
techniques (like linear regression or discriminant analysis)
The main aim this study is to segment Asthma and COPD patients
in to subgroups by their biological (i.e. sputum cytokines)
characteristics
This might help to identify subjects phenotype rather than disease
specific categorisation( which is already labelled as asthma or
COPD)
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

22. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
Cluster analysis is unsupervised statistical techniques which identifies
homogeneous subgroups and creates labels for further supervised
techniques (like linear regression or discriminant analysis)
The main aim this study is to segment Asthma and COPD patients
in to subgroups by their biological (i.e. sputum cytokines)
characteristics
This might help to identify subjects phenotype rather than disease
specific categorisation( which is already labelled as asthma or
COPD)
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

23. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
Cluster analysis is unsupervised statistical techniques which identifies
homogeneous subgroups and creates labels for further supervised
techniques (like linear regression or discriminant analysis)
The main aim this study is to segment Asthma and COPD patients
in to subgroups by their biological (i.e. sputum cytokines)
characteristics
This might help to identify subjects phenotype rather than disease
specific categorisation( which is already labelled as asthma or
COPD)
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

24. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
Cluster analysis is unsupervised statistical techniques which identifies
homogeneous subgroups and creates labels for further supervised
techniques (like linear regression or discriminant analysis)
The main aim this study is to segment Asthma and COPD patients
in to subgroups by their biological (i.e. sputum cytokines)
characteristics
This might help to identify subjects phenotype rather than disease
specific categorisation( which is already labelled as asthma or
COPD)
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

25. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Clustering heterogeneous subjects
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

26. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Clustering heterogeneous subjects
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

27. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Clustering heterogeneous subjects
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

28. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
There are 18 sputum cytokines measurements
Factor analysis was performed on these cytokines, and four
independent factor scores were extracted
The factor scores were used as input variables in K-means cluster
analysis
Thus, three biological clusters was identified
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

29. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
There are 18 sputum cytokines measurements
Factor analysis was performed on these cytokines, and four
independent factor scores were extracted
The factor scores were used as input variables in K-means cluster
analysis
Thus, three biological clusters was identified
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

30. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
There are 18 sputum cytokines measurements
Factor analysis was performed on these cytokines, and four
independent factor scores were extracted
The factor scores were used as input variables in K-means cluster
analysis
Thus, three biological clusters was identified
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

31. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
There are 18 sputum cytokines measurements
Factor analysis was performed on these cytokines, and four
independent factor scores were extracted
The factor scores were used as input variables in K-means cluster
analysis
Thus, three biological clusters was identified
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

32. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Asthma and COPD Biological Clustering
There are 18 sputum cytokines measurements
Factor analysis was performed on these cytokines, and four
independent factor scores were extracted
The factor scores were used as input variables in K-means cluster
analysis
Thus, three biological clusters was identified
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

33. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Three biological Asthma and COPD clusters
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

34. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Pattern of cytokines across the biological clusters
Table 2. Summary statistics across the three identified biological clusters in the test study
Variable Cluster 1 Cluster 2 Cluster 3 C1 vs. C2 C1 vs. C3 C2 vs.C3
(Asthma=55, COPD=3) (Asthma=28, COPD=19) (Asthma=2, COPD=39) P-value P-value P-value
Male [n (%)] 32 (55.2) 26 (55.3) 28 (68.3) 0.99 0.19 0.21
Current or Ex-smokers [n (%)] 22 (37.9) 29 (61.7) 40 (97.6) 0.015 <0.0001 <0.000
Age (years) + 55 (1.5) 60 (2.1) 67 (1.8) 0.038 <0.0001 0.008
Pre FEV1/FVC ratio (%)+ 69.0 (1.9) 58.5 (2.3) 49.7 (2.4) <0.0001 <0.0001 0.011
Pre FEV1 Predicted (%)+ 77.0 (2.7) 59.9 (3.7) 47.0 (3) <0.0001 <0.0001 0.01
Post FEV1 Predicted (%)+ 81.7 (2.7) 63.9 (3.9) 49.1 (3) <0.0001 <0.0001 0.005
Sputum Neutrophil count (%)+ 58.8 (3.1) 77.2 (3) 59.1 (3.1) <0.0001 0.95 <0.0001
Sputum Eosinophil count (%) 3.9 (2.4 - 6.4) 0.7 (0.5 - 0.9) 2.0 (1.25 - 3.17) <0.0001 0.039 <0.0001
CFU >107/ml or positive culture (n[%]) 8 (13.8) 26(55.3) 9 (21.9) <0.0001 0.29 0.001
IL-1 (pg/ml) 39.5 (30.8 - 50.8) 379.5 (257.3 - 559.8) 23.5 (17.2 - 32.2) <0.0001 0.025 <0.0001
IL-5 (pg/ml) 2.6 (1.6 - 4.2) 2.2 (1.4 - 3.4) 1.4 (0.9 - 2.2) 0.56 0.083 0.22
IL-6 (pg/ml) 21.3 (15 - 30.4) 271.4 (192.2 - 383.3) 486.2 (327.7 - 721.4) <0.0001 <0.0001 0.031
IL-6R (pg/ml) 163.2 (126.0 - 211.6) 433.4 (344.2 - 545.6) 112.4 (88.6 - 142.6) <0.0001 0.04 <0.0001
IL-10 (pg/ml) 0.33 (0.25 - 0.45) 5.5 (3.5 - 8.7) 0.34 (0.2 - 0.6) <0.0001 0.89 <0.0001
IL-13 (pg/ml) 10.4 (7.7 - 14.0) 4.8 (3.8 - 6.2) 3.5 (2.4 - 5.2) 0.001 <0.0001 0.18
CCL-26 (pg/ml) 12.4 (7.8 - 19.9) 5.0 (3.4 - 7.5) 2.9 (1.9 - 4.6) 0.004 <0.0001 0.081
TNF↵ (pg/ml) 1.4 (1. 1 - 1.9) 29.9 (19.5 - 45.9) 1.7 (1.1 - 2.5) <0.0001 0.62 <0.0001
Data presented as Geometric mean with corresponding confidence interval unless stated, +
Mean (SEM) ; CFU= colony forming units;C=cluster
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

35. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Table of contents
1 Introduction and Objectives
2 Asthma and COPD Biological Clustering
3 Asthma and COPD Validation Study
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
4 Summary and Conclusion
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

36. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation Study
This study was validated on independent severe asthma and COPD
studies.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

37. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation Study
This study was validated on independent severe asthma and COPD
studies.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

38. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Asthma and COPD Validation Study
For validation, independent severe asthma (n=166) and COPD
(n=58) patients were used
Two classification techniques were applied; i.e.
1. Linear discriminant analysis (LDA)
2. The diseases status (Asthma or COPD), and IL-1 cuto↵ for the
overlap
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

39. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Asthma and COPD Validation Study
For validation, independent severe asthma (n=166) and COPD
(n=58) patients were used
Two classification techniques were applied; i.e.
1. Linear discriminant analysis (LDA)
2. The diseases status (Asthma or COPD), and IL-1 cuto↵ for the
overlap
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

40. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Asthma and COPD Validation Study
For validation, independent severe asthma (n=166) and COPD
(n=58) patients were used
Two classification techniques were applied; i.e.
1. Linear discriminant analysis (LDA)
2. The diseases status (Asthma or COPD), and IL-1 cuto↵ for the
overlap
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

41. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Asthma and COPD Validation Study
For validation, independent severe asthma (n=166) and COPD
(n=58) patients were used
Two classification techniques were applied; i.e.
1. Linear discriminant analysis (LDA)
2. The diseases status (Asthma or COPD), and IL-1 cuto↵ for the
overlap
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

42. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Asthma and COPD Validation Study
For validation, independent severe asthma (n=166) and COPD
(n=58) patients were used
Two classification techniques were applied; i.e.
1. Linear discriminant analysis (LDA)
2. The diseases status (Asthma or COPD), and IL-1 cuto↵ for the
overlap
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

43. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Characteristics of the Validation study
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

44. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Table of contents
1 Introduction and Objectives
2 Asthma and COPD Biological Clustering
3 Asthma and COPD Validation Study
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
4 Summary and Conclusion
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

45. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validating Asthma and COPD using LDA
Classification model was developed from the Test study using LDA
Then subjects were assigned to subgroups in which he/she has the
highest discriminant score/posterior probability.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

46. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validating Asthma and COPD using LDA
Classification model was developed from the Test study using LDA
Then subjects were assigned to subgroups in which he/she has the
highest discriminant score/posterior probability.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

47. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validating Asthma and COPD using LDA
Classification model was developed from the Test study using LDA
Then subjects were assigned to subgroups in which he/she has the
highest discriminant score/posterior probability.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

48. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validating Asthma and COPD using LDA
Classification model was developed from the Test study using LDA
Then subjects were assigned to subgroups in which he/she has the
highest discriminant score/posterior probability.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

49. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validating Asthma and COPD using LDA
Table 3. Coe cients (betas) and prior probability of each cluster in the test
study which used to predict class membership in the validation study
Variables Cluster 1 ( 1k ) Cluster 2 ( 2k ) Cluster 3 ( 3k )
IL-1 1.63 2.17 0.96
IL-5 -6.31 -6.50 -6.79
IL-6 -2.62 -1.17 0.14
IL-6R 6.28 5.58 4.72
IL-8 6.64 7.59 7.31
CCL-2 5.56 5.66 6.98
CCL-4 3.59 3.83 4.66
CCL-5 -1.36 -0.80 -2.83
CCL-13 -0.04 -0.49 -0.21
CCL-17 2.13 1.09 1.09
CXCL-11 0.48 0.51 0.29
TNF↵ -5.77 -5.28 -6.57
Constant -63.39 -75.75 -79.11
Prior Probability 0.33 0.33 0.33
Unit of all cytokines is pg/ml
Sij = j + j1Y1i + j2Y2i + · · · + jk Yki + log(Pj )
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

50. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Patterns of characteristics across validation subgroups
Table 4. Summaries across the validation subgroups which were identified using linear discriminant analysis (LDA)
Variable Group 1 Group 2 Group 3 G1 vs. G2 G1 vs. G3 G2 vs.G3
(Asthma=94, COPD=12) (Asthma=55, COPD=18) (Asthma=7, COPD=28) P-value P-value P-value
Male [n (%)] 65 (61.3) 40 (54.8) 25 (71.4) 0.38 0.28 0.1
Current or Ex-smokers 37 (34.9) 42 (57.5) 33 (94.3) 0.003 <0.0001 <0.0001
Age (year)+ 53 (1.3) 56 (2.0) 66 (2.0) 0.15 <0.0001 0.002
Pre FEV1/FVC ratio (%)+ 68.2 (1.4) 61.8 (1.9) 57.3 (1.8) 0.004 <0.0001 0.14
Pre FEV1 Predicted (%)+ 67.6 (2.2) 64.7 (2.9) 65.5 (3.2) 0.4 0.65 0.85
Sputum Neutrophil count (%)+ 59.1 (2.8) 72.3 (2.5) 58.4 (4.0) 0.001 0.88 0.003
Sputum Eosinophil count (%) 6.3 (4.9 - 8.2) 3.0 (2.4 - 3.8) 3.2 (2.3 - 4.4) <0.0001 0.003 0.79
IL-1 (pg/ml) 54.1 (42.7 - 68.5) 526.6 (375.5 - 738.4) 42.4 (26 - 69.2) <0.0001 0.36 <0.0001
IL-5 (pg/ml) 1.4 (1.1 - 1.8) 1.2 (0.8 - 1.7) 0.9 (0.5 - 1.5) 0.43 0.085 0.34
IL-6 (pg/ml) 26.1 (19.9 - 34.3) 273.3 (210.6 - 354.6) 344.2 (237.2 - 499.3) <0.0001 <0.0001 0.32
IL-6R (pg/ml) 186.0 (135.5 - 255.3) 589.8 (476.6 - 729.8) 90.5 (50.3 - 162.8) <0.0001 0.013 <0.0001
IL-10 (pg/ml) 1.9 (1.5 - 2.4) 3.9 (2.8 - 5.4) 0.5 (0.3 - 0.9) 0.001 <0.0001 <0.0001
TNF↵ (pg/ml) 1.4 (1.2 - 1.7) 23.3(16.1 - 33.7) 1.8 (1.1 - 3.0) ¡0.0001 0.32 <0.0001
Data presented as geometric mean with corresponding confidence interval unless stated, +
Mean (SEM); G=Group
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

51. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Cytokines profiles in the test and validation subgroups
Figure : Test Figure : Validation
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

52. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation Results: Using LDA
Validation Results
The sputum cellular and cytokines patterns were very similar between the
test and validation subgroups.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

53. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validating Asthma and COPD using LDA
Table 3. Coe cients (betas) and prior probability of each cluster in the test
study which used to predict class membership in the validation study
Variables Cluster 1 ( 1k ) Cluster 2 ( 2k ) Cluster 3 ( 3k )
IL-1 1.63 2.17 0.96
IL-5 -6.31 -6.50 -6.79
IL-6 -2.62 -1.17 0.14
IL-6R 6.28 5.58 4.72
IL-8 6.64 7.59 7.31
CCL-2 5.56 5.66 6.98
CCL-4 3.59 3.83 4.66
CCL-5 -1.36 -0.80 -2.83
CCL-13 -0.04 -0.49 -0.21
CCL-17 2.13 1.09 1.09
CXCL-11 0.48 0.51 0.29
TNF↵ -5.77 -5.28 -6.57
Constant -63.39 -75.75 -79.11
Prior Probability 0.33 0.33 0.33
Unit of all cytokines is pg/ml
Sij = j + j1Y1i + j2Y2i + · · · + jk Yki + log(Pj )
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

54. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Table of contents
1 Introduction and Objectives
2 Asthma and COPD Biological Clustering
3 Asthma and COPD Validation Study
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
4 Summary and Conclusion
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

55. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
IL-1 across the three clusters
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

56. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
IL-1 across the three clusters at 130 pg/ml
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

57. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation using IL-1 cuto↵ for the overlap and diseases
Since IL-1 performed well in discriminating the overlap group in the
test study at 130 pg/ml cuto↵
This cuto↵ point was applied to the validation studies
With assumption that asthma and COPD are clinically
distinguishable diseases
Then the validation dataset classified in to three subgroups
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

58. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation using IL-1 cuto↵ for the overlap and diseases
Since IL-1 performed well in discriminating the overlap group in the
test study at 130 pg/ml cuto↵
This cuto↵ point was applied to the validation studies
With assumption that asthma and COPD are clinically
distinguishable diseases
Then the validation dataset classified in to three subgroups
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

59. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation using IL-1 cuto↵ for the overlap and diseases
Since IL-1 performed well in discriminating the overlap group in the
test study at 130 pg/ml cuto↵
This cuto↵ point was applied to the validation studies
With assumption that asthma and COPD are clinically
distinguishable diseases
Then the validation dataset classified in to three subgroups
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

60. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation using IL-1 cuto↵ for the overlap and diseases
Since IL-1 performed well in discriminating the overlap group in the
test study at 130 pg/ml cuto↵
This cuto↵ point was applied to the validation studies
With assumption that asthma and COPD are clinically
distinguishable diseases
Then the validation dataset classified in to three subgroups
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

61. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation using IL-1 cuto↵ for the overlap and diseases
Since IL-1 performed well in discriminating the overlap group in the
test study at 130 pg/ml cuto↵
This cuto↵ point was applied to the validation studies
With assumption that asthma and COPD are clinically
distinguishable diseases
Then the validation dataset classified in to three subgroups
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

62. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Patterns of characteristics across the groups in validation
Table 5. Summaries across the validation subgroups which were identified using IL-1 cuto↵ and diseases status (asthma or COPD)
Variable Group 1 Group 2 Group 3 G1 vs. G2 G1 vs. G3 G2 vs.G3
(Asthma=103) (Asthma=63, COPD=26) (COPD=32) P-value P-value P-value
Male [n (%)] 66 (64.1) 51 (57.3) 22 (68.7) 0.34 0.63 0.26
Current or Ex-smokers 28 (27.2) 54 (60.7) 32 (100) <0.0001 <0.0001 <0.0001
Age (year)+ 49 (1.2) 57 (1.8) 68 (1.6) <0.0001 <0.0001 0.001
Pre FEV1/FVC ratio (%)+ 70.1 (1.2) 61.0 (1.7) 56.6 (2.4) <0.0001 <0.0001 0.16
Pre FEV1 Predicted (%)+ 71.2 (2.3) 64.1 (2.5) 60.2 (3.6) 0.033 0.019 0.4
Sputum Neutrophil count (%)+ 59.0 (2.8) 70.4 (2.5) 59.2 (4.2) 0.003 0.97 0.023
Sputum Eosinophil count (%) 5.6 (4.3 - 7.3) 3.5 (2.8 - 4.4) 3.9 (2.5 - 5.9) 0.009 0.12 0.68
IL-1 (pg/ml) 37.0 (29.1 - 47) 527.1(407.1 - 682.5) 40.0 (28.2 - 56.6) <0.0001 0.75 <0.0001
IL-5 (pg/ml) 1.2 (1.0 - 1.5) 1.3 (1.0 - 1.9) 1.0 (0.6 - 1.7) 0.69 0.45 0.37
IL-6 (pg/ml) 34.7 (25.6 - 47) 190 (138.4 - 261) 157.7 (88.8 - 280.2) <0.0001 <0.0001 0.56
IL-6R (pg/ml) 153(102.7 - 228.6) 549.4 (454.9 - 663.6) 101.7(74.1-139.5) <0.0001 0.17 <0.0001
IL-10 (pg/ml) 2.2 (1.8 - 2.6) 3.1 (2.2 - 4.3) 0.4 (0.2 - 0.6) 0.063 < 0.0001 <0.0001
TNF- (pg/ml) 1.6 (1.3 - 2.0) 14.4 (9.8 - 21.2) 1.3 (0.7 - 2.1) <0.0001 0.39 <0.0001
Data presented as geometric mean with corresponding confidence interval unless stated, +
Mean (SEM) ; CFU= colony forming units; G=Group
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

63. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Cytokines profiles in the test and validation subgroups
Figure : Test Figure : Validation ( using IL-1 )
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

64. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Validation Results: Using diseases and IL-1 cuto↵
Validation Results
The sputum cellular and cytokines patterns were also very similar
between the test and validation subgroups.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

65. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
Cytokine profiles in the test and validation by clusters
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

66. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
TNF↵ as alternative for IL-1
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

67. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
TNF↵ as alternative for IL-1
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

68. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
TNF↵ as alternative for IL-1
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

69. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Table of contents
1 Introduction and Objectives
2 Asthma and COPD Biological Clustering
3 Asthma and COPD Validation Study
Validating Asthma and COPD using LDA
Validating Asthma and COPD using IL-1 cuto↵
4 Summary and Conclusion
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

70. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
Three biological clusters were identified which are...
Cluster 1
Eosinophilic asthma predominant (95% asthma) high Th-2 cytokines
Cluster 2
Neutrophilic asthma and COPD overlap representing about one-third of
each disease group, high in Th-1 and proinflmmatory(PI) cytokines
Cluster 3
Non-neutrophilic COPD predominant (95% COPD) and high in PI
cytokines
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

71. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
Three biological clusters were identified which are...
Cluster 1
Eosinophilic asthma predominant (95% asthma) high Th-2 cytokines
Cluster 2
Neutrophilic asthma and COPD overlap representing about one-third of
each disease group, high in Th-1 and proinflmmatory(PI) cytokines
Cluster 3
Non-neutrophilic COPD predominant (95% COPD) and high in PI
cytokines
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

72. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
Three biological clusters were identified which are...
Cluster 1
Eosinophilic asthma predominant (95% asthma) high Th-2 cytokines
Cluster 2
Neutrophilic asthma and COPD overlap representing about one-third of
each disease group, high in Th-1 and proinflmmatory(PI) cytokines
Cluster 3
Non-neutrophilic COPD predominant (95% COPD) and high in PI
cytokines
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

73. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
Three biological clusters were identified which are...
Cluster 1
Eosinophilic asthma predominant (95% asthma) high Th-2 cytokines
Cluster 2
Neutrophilic asthma and COPD overlap representing about one-third of
each disease group, high in Th-1 and proinflmmatory(PI) cytokines
Cluster 3
Non-neutrophilic COPD predominant (95% COPD) and high in PI
cytokines
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

74. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
Three biological clusters were identified which are...
Cluster 1
Eosinophilic asthma predominant (95% asthma) high Th-2 cytokines
Cluster 2
Neutrophilic asthma and COPD overlap representing about one-third of
each disease group, high in Th-1 and proinflmmatory(PI) cytokines
Cluster 3
Non-neutrophilic COPD predominant (95% COPD) and high in PI
cytokines
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

75. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
This study was validated on independent study using two methods,
and reveal very similar pattern in cytokines and sputum cellular
IL-1 (at 130 pg/ml) and TNF↵ (at 5 pg/ml) preformed reasonably
well in discriminating the overlap subgroup, and could be potential
future markers
Conclusion
In conclusion, sputum cytokines can determine distinct and overlapping
groups of patients with severe asthma and COPD, and may contribute to
improved patients classification to enable stratified medicine!
However,for generalisation this findings need to be replicate in
multi-studies
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

76. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
This study was validated on independent study using two methods,
and reveal very similar pattern in cytokines and sputum cellular
IL-1 (at 130 pg/ml) and TNF↵ (at 5 pg/ml) preformed reasonably
well in discriminating the overlap subgroup, and could be potential
future markers
Conclusion
In conclusion, sputum cytokines can determine distinct and overlapping
groups of patients with severe asthma and COPD, and may contribute to
improved patients classification to enable stratified medicine!
However,for generalisation this findings need to be replicate in
multi-studies
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

77. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
This study was validated on independent study using two methods,
and reveal very similar pattern in cytokines and sputum cellular
IL-1 (at 130 pg/ml) and TNF↵ (at 5 pg/ml) preformed reasonably
well in discriminating the overlap subgroup, and could be potential
future markers
Conclusion
In conclusion, sputum cytokines can determine distinct and overlapping
groups of patients with severe asthma and COPD, and may contribute to
improved patients classification to enable stratified medicine!
However,for generalisation this findings need to be replicate in
multi-studies
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

78. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
This study was validated on independent study using two methods,
and reveal very similar pattern in cytokines and sputum cellular
IL-1 (at 130 pg/ml) and TNF↵ (at 5 pg/ml) preformed reasonably
well in discriminating the overlap subgroup, and could be potential
future markers
Conclusion
In conclusion, sputum cytokines can determine distinct and overlapping
groups of patients with severe asthma and COPD, and may contribute to
improved patients classification to enable stratified medicine!
However,for generalisation this findings need to be replicate in
multi-studies
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

79. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Summary and Conclusion
This study was validated on independent study using two methods,
and reveal very similar pattern in cytokines and sputum cellular
IL-1 (at 130 pg/ml) and TNF↵ (at 5 pg/ml) preformed reasonably
well in discriminating the overlap subgroup, and could be potential
future markers
Conclusion
In conclusion, sputum cytokines can determine distinct and overlapping
groups of patients with severe asthma and COPD, and may contribute to
improved patients classification to enable stratified medicine!
However,for generalisation this findings need to be replicate in
multi-studies
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

80. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
References
Carolan BJ and Sutherland ER (2013); Clinical phenotypes of chronic obstructive
pulmonary disease and asthma: Recent advances.
Tabachnick BG and Fidell LS (2007); Using Multivariate Statistics. FIFTH ed.
Everitt B (2011); An Introduction to Applied Multivariate Analysis with R.
Gregory R and Samuelsen KM (2008); Advances in Latent Variable Mixture
Models.
Barnes PJ (2008); The cytokine network in asthma and chronic COPD disease.
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma

81. Introduction and Objectives
Asthma and COPD Biological Clustering
Asthma and COPD Validation Study
Summary and Conclusion
References
Acknowledgement
Prof Chris Brightling, Prof Paul Burton & Dr Chris Newby
Dr Mona Bafadhel & Dr Dhan Desai
Michael A Ghebre University of Leicester Biological clustering supports both Dutch and British hypotheses of Asthma