Investigation Of Biomarker For Prediction Of Response To Treatment In Chronic Obstructive Pulmonary Disease

Background: Chronic Obstructive Pulmonary Disease (COPD) is a disease state characterized by airflow limitation that is not fully reversible. The pathological change not only limited to the pulmonary , but also has the systemic(also called extrapulmonary) effect. The main purpose of treatment to COPD is to improve the symptom, improve the quality of life and lung function, reduce exacerbation and mortality. It is not clear that, however, whether the COPD patients respond the same to treatment? Whether COPD patients with different clinic characteristic have the different respondence to treatment? Whether the different expression of biomarker(s) in COPD patients affect the therapy respondence? And whether the biomarker(s) of COPD have the predicted effect of therapy respondence to treatment?Purpose: To explore the predicted effect of biomarker in COPD to treatment, and to distinguish the subgroup of COPD patients that have the better therapy respondence, finally to accomplish the goal of personally treatment.Materials and methods:1. Research Object: Sixty-five stable COPD patients and 26 gender and age-matched healthy volunteers were recruited for the study. Patients were recruited from outpatients department of Guangzhou Institute of Respiratory Disease, the first affiliated hospital of Guangzhou Medicial Colloge during the period of Mar 2009 to Feb 2010. All COPD subjects were at the stable stage,and COPD was diagnosed according to the Global Initiative for Chronic Obstructive Lung Disease guidelines from observing symptoms of progressive shortness of breath, productive cough, and occasional wheezing. Airflow limitation was characterized by FEV1/FVC ratio<70% after the inhalation of a bronchodilator. All subjects were nonatopic, had no history of rhinitis, eczema, or asthma, and had been free from respiratory tract infections or exacerbations and had no use of oral or inhaled corticosteroid for 4 weeks prior to undergoing of pulmonary function test and sputum induction. Subjects in control group were recruited from health examination department of the first affiliated hospital of Guangzhou Medical College,these subjects had the normal pulmonary function test and had been free from respiratory tract infections for 4 weeks.2. Methods: The study participants of COPD patients received Fluticasone Propionate (250μg twice daily) and Long-Acting Beta2-Agonists Salmeterol (50μg twice daily) for 4 weeks. All other medications, including short-acting Beta2-adrenoceptor agonists, anticholinergics, and antibiotics, were permitted during the study. At the beginning of the study and 1 month later, the biomarkers(Matrix Metalloproteinase-9,Tissue Inhibitor of Metalloproteinase-1,desmosine,Pulmonary Surfactant Protein D,Clara Cell Secreted Protein-16) in serum and induced sputum supernatant,pulmonary function including FEV1,FVC,FEV1/FVC and MRC dyspnea scal,St.George's respiratory Questionnaires(SGRQ) were collected; according to the expression of biomarkers of serum and sputum, COPD patients were divided into two grpups respectively: the high express group and the low express group, and the change of FEV1,FVC,MRC,SGRQ score after 1 month treatment were compared.The multiple regression analysis and logistic regression analysis were conducted for indexs that related to the therapy respondence.Results:1. Baseline demographic and clinical characteristics There were 65 COPD patients in this study: 61 men and 4 women, average age(y) 66.6±8.1; and 26 normal control: 22 men and 4 women, aged 66.7±10, there was no significant difference of gender constituent ratio and average age between the two groups(P>0.05); The body mass index (kg/m2) of the two groups had no significant difference (21.8±3.42 Vs 22.5±2.8, P>0.05); The smoking index(pack-year) of COPD group(41.7±19.7) was higher than normal control group (8.96±15.6); The FEV1 (L),FEV1%pre(%),FVC(L),FEV1/FVC(%) of COPD group were: 1.08±0.54,42.93±18.14,2.33±0.76,45.65±12.13, lower than normal control group significantly (P<0.001, respectively). 2.The change of pulmonary function,MRC,SGRQand biomarkersAfter 1 month treatment, the pulmonary function,MRC,SGRQ score had been significantly improved (FEV1(L): 103±0.46 to 1.15±0.48, FVC(L): 2.32±0.74 to 2.51±0.60),MRC score(2.49±1.22 to 1.59±1.19),SGRQ score(52.62±17.47 to 35.38±18.10)(P < 0.001,respectively);the serum TIMP-1 concentration decreased significantly(595.13±163.03 to 300.55±166.40 ng/ml)and serum CC-16 concentration improved significantly(5.94±1.77 to 7.22±1.87 ng/ml)(t value 3.352,-4.292, P value 0.003,<0.001, respectively); the induced sputum cytology categorization and other biomarkers in serum and sputum supernatant, however, had no significant change(P>0.05,respectively).3. Bronchodilator reversibility and therapy respondence3.1 There were 65 COPD patients recruited in the study, 63 of them finished the bronchodilator reversibility test, and 10 of 63 patients (15.87%) had bronchodilator reversibility; 53 of 63 patients(84.13%) had no bronchodilator reversibility.3.2 According to the bronchodilator reversibility test, COPD patients were divided into two groups: positively group and negatively group. The clinical characteristic of the two groups were compared and found: the disease course of positively group was shorter (5.67±2.95 Vs 8.92±7.04, t=-3.563, P =0.001), and the FEV1,FVC value were higher than the negatively group(FEV1:1.15±0.33 Vs 0.96±0.46, FVC:2.88±0.60 Vs 2.30±0.71; t value 2.032,3.863; P value 0.044,<0.001, respectively);Other indexs such as gender constituent ratio,age,BMI,MRC,SGRQ score and airway inflammatory characteristic (sputum cytology categorization) had no significant difference between the two groups(P>0.05, respectively).3.3 COPD bronchodilator reversibility and the therapy respondenceAfter 1 month treatment , the change of pulmonary function,MRC,SGRQ were compared between positively group and negatively group, and founded: the change of SGRQ total score and impact score in positively group(27.00±16.92, 29.00±23.13, respectively) were higher than negatively group(14.52±12.56,12.92±13.28, respectively) significantly (t value2.047,2.292;P value 0.050,0.029) , the change of other indexs had no significant difference between the two groups(P>0.05 ,respectively).4. Induced sputum and COPD4.1 There were 57 COPD patients finished the sputum induction, and 51 of them got qualified sputum; 22 normal control subjects finished the sputum induction, 18 of them got qualified sputum. The mean sputum weight(g) of COPD group(0.51±0.08) was higher than normal control group(0.46±0.13);the neutrophil(Neu),macrophage (Mac),eosinophil(Eos),lymphocyte(Lym) constituent ratio(%)were: 86.24±15.04,5.75±6.96,4.71±4.79,1.30±1.09 in COPD group, and 88.41±10.99,9.59±11.13,1.26±1.20,0.74±0.56 in normal control group, there had no significant difference between the former two(t value -0.547,-1.336,P value 0.586,0.196,respectively) ,but the difference between the later two is significantly(t value 4.677,2.747, P value <0.001, 0.008 , respectively). Also, the subjects with higher sputum Eos (Eos≥2.5%) in COPD group was 60.78% and 0% in normal control group(F=19.867, P<0.001).4.2 Sputum Eos and the COPD severitySputum Eos was inversely associated with FEV1/FVC ratio (r=-0.399, P=0.004) , and sputum Eos had no association with other indexs (P>0.05, respectively).4.3 Sputum Eos and the therapy respondenceAccording to the sputum Eos, COPD patients were divided into two groups: the high Eos group (Eos≥2.5%) and the low Eos group (Eos<2.5%). The change of pulmonary function,MRC,SGRQ score were compared between the two groups, and found: the change of FEV1(L) and SGRQ symptom score in the high Eos group (0.16±0.18, 21.50±17.24, respectively) were higher than the low Eos group (0.05±0.09, 10.93±9.94, respectively) (t value 2.603,2.331, P value 0.014,0.026, respectively) , the change of other indexs had no significant difference between the two groups(P>0.05, respectively).5. Matrix Metalloproteinase-9 (MMP-9),Tissue Inhibitor of Metalloproteinase -1 (TIMP-1) and their ratio and COPD5.1 The expression of MMP-9,TIMP-1 and their ratioIn stable COPD, the serum MMP-9(318.24±158.22ng/ml),TIMP-1(493.64±116.77 ng/ml) were higher than normal control (255.93±104.66ng/ml, 422.30±73.73ng/ml, respectively)(t value 2.103,3.019, P value 0.039,0.003, respectively); MMP-9/ TIMP-1 ratio had no significant difference between the two groups(t=0.45, P=0.654).5.2 Serum MMP-9,TIMP-1,MMP-9/TIMP-1 ratio and COPD severitySerum MMP-9,TIMP-1,MMP-9/TIMP-1 ratio had no association with pulmonary function (P>0.05, respectively); TIMP-1 was positively associated with MRC and SGRQ total score,activity score,impact score(r value 0.384,0.384,0.272,0.425, P value 0.005,0.005,0.049,0.002, respectively)。5.3 COPD serum MMP-9,TIMP-1,MMP-9/TIMP-1 ratio and therapy respondenceCOPD patients were divided into two groups according to the serum MMP-9,TIMP-1,MMP-9/TIMP-1 ratio:the high express group and the low express group, the change of pulmonary function,MRC,SGRQ were compared, and found: the change of FVC in low MMP-9 and MMP-9/TIMP-1 ratio express group(FVC(ml) change:318±407, 346±453, respectively) was higher than high express group( FVC(ml) change :41±197,97±246, respectively) (P<0.05 ,respectively); the change of FEV1(ml),MRC in low TIMP-1 group(137±189 1.00±1.00, respectively)was higher than high express group(83±103,0.64±1.01, respectively)(P<0.05). MMP-9,TIMP-1,MMP-9/TIMP-1 ratio expression had no association with the SGRQ change (P>0.05, respectively).6. Desmosine and COPD6.1 The sputum supernatant desmosine in COPD group was higher than normal control(1062.2±945.22ng/ml Vs 443.49±501.72ng/ml, t=2.277,P=0.027); while the serum desmosine had no significant difference between the two groups(t=0.636, P=0.0527).6.2 Desmosine expression and COPD severityThe serum desmosine had no association with pulmonary function (P >0.05,respectively); but the sputum supernatant desmosine in COPD group was inversely associated with FEV1 and FVC(r value -0.357,-0.479, P value 0.02,0.001,respectively),and had no association with FEV1%pre and FEV1/FVC (P>0.05, respectively). The desmosine in serum or sputum supernatant had no association with MRC and SGRQ score(P>0.05, respectively).6.3 Desmosine express and therapy respondenceAccording to the desmosine express,COPD patients were divided into two groups: the high express group and the low express group. The change of pulmonary function,MRC,SGRQ were compared,and found: the serum desmosine had no association with the change of pulmonary function,MRC,SGRQ(P>0.05, respectively); while the change of MRC was higher in low sputum desmosine group(1.24±1.15) than high express group(0.40±0.52,t=-2.589,P=0.016) , the change of other indexs were higher in low sputum desmosine express group, but had no statistical difference(P>0.05, respectively).7.Surfactant protein D(SP-D) and COPD7.1 The serum and sputum supernatant SP-D in COPD group was higher than the normal control(serum: 45.46±37.78 Vs 31.68±12.04 ng/ml; sputum: 173.23±186.93 Vs 89.59±70.29ng/ml, t value 2.236,2.578, P value 0.021,0.012, respectively)。7.2 SP-D and COPD severity The serum and sputum supernatant SP-D express in COPD group had no association with pulmonary function,MRC,SGRQ score(P均>0.05, respectively).7.3 SP-D express and therapy respondence According to the SP-D express, COPD patients were divided into two groups:the high express group and the low express group. The change of pulmonary function,MRC,SGRQ were compared,and found that: the change of pulmonary function,MRC,SGRQ score had no significant difference between the two groups (P > 0.05, respectively).8. Clara cell secreted protein-16(CC-16) and COPD8.1 The serum CC-16 express in COPD group (6.20±2.7ng/ml) was lower than the normal control(7.83±2.15ng/ml, t=-2.752, P=0.007); while there is no significant difference of sputum CC-16 between the two groups(t=1.460, P=0.149).8.2 CC-16 and COPD severityThe serum and sputum supernatant CC-16 express in COPD group had no association with pulmonary function,MRC,SGRQ score(P>0.05, respectively).8.3 CC-16 express and COPD therapy respondenceAccording to the CC-16 express, COPD patients were divided into two groups:the high express group and the low express group. The change of pulmonary function,MRC,SGRQ were compared, and found that: the had no significant difference of pulmonary function,MRC,SGRQ score change between the two groups (P>0.05, respectively);while the change of serum CC-16 concentration had the association with MRC improvement.9. Multiple regression analysis result: the basic FVC%pre, MMP-9/TIMP-1 and sputum CC-16 concentration negatively associated with FVC change; smoking index, basic MRC positively and basic sputum desmosine negatively associated with MRC change; smoking index, basic impact score positively associated with SGRQ change; smoking index, basic activity score positively associated with activity change;smoking index, basic activity score positively and basic FVC negatively associated with impact change; the basic symptom positively associated with symptom change (P<0.05, respectively);and there had no index associated with FEV1 change.10. Logistic regression analysis result: The basic FEV1,FVC have the association with theray respondence (FEV1 change≥5% and/or FVC change≥10%) .Conclusion:The present study explored the change of pulmonary function, MRC, SGRQ score after 1 month of treatment, and also the pathophysiological index that related to the change were also explored in mild to very severe COPD patients, the main results are listed as follow:1. There were15.87% of COPD patients had bronchodilator reversibility, although these patients had a better change of SGRQ total score and impact score, the bronchodilator reversibility had no predicted effect of pulmonary function change in these patients.2. There were 60.78% COPD patients had induced sputum eosinophil; the high sputum eosinophil was correlated significantly with the airflow limitation and these patients had a better therapy respondence to treatment. 3. The serum MMP-9,TIMP-1 of COPD patients were higher than normal control. The high express of TIMP-1 was correlated significantly with MRC and SGRQ of these patients; the express of MMP-9,TIMP-1,MMP-9/TIMP-1 were correlated with the therapy respondencd: the lower expression, the better respondence.4. The serum and sputum supernatant SP-D of COPD patients were higher than normal control. However, the high SP-D had no relationship with the pulmonary function,MRC,SGRQ and had no relationship with the therapy respondence.5. The serum CC-16 of COPD patients were lower than normal control. CC-16 reflex the respiratory epithelial cells function, and the change of serum CC-16 concentration had the association with MRC improvement.6. With regard to COPD patients, the smoking index, basic MRC score positively and sputum desmosine negatively associated with MRC change; smoking index had the positive association with symptom change too.7. The basic FEV1,FVC had the association with treatment respondence (FEV1 change ratio≥5% and/or FVC change ratio≥10%).