proteomic evaluation of Chikungunya
- Conditions
- Antibody positive and negative Healthy volunteersChikungunya virus disease,
- Registration Number
- CTRI/2019/03/018051
- Lead Sponsor
- Research Society
- Brief Summary
**Background and Rationale:**
Chikungunyais a viral disease transmitted by Aedes mosquitoes. Chikungunya virus (CHIKV)is an arbovirus that belongs to the family of Togaviridae and genus Alphavirus.The virus was first discovered in 1952 and used tocause sporadic outbreaks.
Inrecent years, there have been several reports of large scale outbreaks ofChikungunya virus infection in several parts of Southern India.Over,2000 cases of Chikungunya fever have also been reported from Malegaon town inNasik district, Maharashtra state, India between February-March 2006.In October 2009, 2–3 months after an outbreak of a febriledisease with joint pain on the eastern coast of Madagascar, serologic markersfor chikungunya virus (CHIKV) in n= 1,244 pregnant women were assessed and IgGsero-prevalence against CHIKV was 45%. The recent outbreaks of this diseasehave been associated with fatalities and chronic and persistent disabilityparticularly the persistence of joint pain. Chikungunya is thus considered asan important re-emerging public health problem in both tropicaland temperate countries, where the distribution of the Aedes mosquito vectorscontinues to expand.
Giventhis backdrop, several pharmaceutical companies are in the process ofdeveloping vaccines to combat the disease. One such vaccine has been developedby Bharat Biotech and is undergoing clinical development at the KEM Hospital. APhase I study [a study in normal, healthy participants is currently underway atthis institute after regulatory and Ethics Committee approval- EC-PHARMA-10/16].
Whilescreening normal, healthy participants for inclusion into this study the studyteam found 13/53 [25%] participants to be positive for anti- Chikungunyaantibodies by an ELISA test. When questioned, these 13 participants did notgive any past history of Chikungunya infection indicating that theysuffered from asymptomatic infection.When the team did aliterature review of the prevalence of anti-Chikungunya antibodies in thegeneral population, there were no studies. Only three studies on antibodyprevalence have been done in the country and these are largely confined topatients who have been hospitalized with a diagnosis of Chikungunya.
A diseaselike Chikungunya that has seen a re-emergence would benefit from a biomarkerthat facilitates its diagnosis and helps assess prognosis. Biomarker studiesbeing with an assessment of plasma proteins that are up or down regulatedduring and after the infection – an analysis that can be done by proteomics- astudy of an entire set of proteins that are modified by an organism or system.
Thus,the present study is planned with the objective of carrying out a quantitativeproteomic analysis of patients who are positive foranti-Chikungunya antibodies including those from the above-mentioned study(EC/PHARMA-10/16) as well as those who have active infection and analyseproteins that are upregulated or downregulated in them.
**Objective:**
Toassess via a quantitative proteomic analysis the differential expression ofproteins in Chikungunya antibody positive patients (symptomatic andasymptomatic) versusage and sex matched healthycontrols.
**Methods:**
*Ethics*:The study will be submitted to the Institutional EthicsCommittee for approval. Written, informed consent will be taken from allparticipants. A copy of the submissionform will be submitted to the Research Society
*Study design*:Cross sectional
SampleSize calculation: No formal sample size calculation
**Adults:**The department is currently doing a studyEC/PHARMA-10/2016 where a new Chikungunya vaccine is being tested in normal,healthy participants. A total of n = 25 participants who are asymptomatic buttested positive (>11 standard units), n = 25 who satisfy the confirmed casedefinition of Chikungunya (NVBDCP) and n = 50 controls who have tested negative(<11 standard units) will be enrolled in this cross-sectional study.
**Paediatrics:**A total of n = 10 participants who are asymptomatic but tested positive (>11standard units), n = 10 who satisfy the confirmed case definition ofChikungunya (NVBDCP) and n = 20 controls who have tested negative (<11standard units) will be enrolled in this cross-sectional study.
*Screening*:All participants will be screened (via medical history and physical examinationonly) to establish the eligibility criteria as outlined below. This includesnegative antibodies, CHIKV IgG, by ELISA method.
*Selection criteria***:**
**Adults:**Allasymptomatic participants from the Chikungunya vaccine trial (EC/PHARMA-10/16),aged ≥18 and ≤50 years and who are anti-CHIKV antibody positive (>11standard units) will be enrolled as asymptomatic adult cases. These are theparticipants who have been excluded from the vaccine study due to antibody positivity.Symptomatic confirmed adult cases (NVBDCP) will be enrolled from the Departmentof Medicine. Age and sex matched adult controls will be taken from thoserelatives who accompany patients to the Therapeutic Drug Monitoring OPD run bythe Department on Fridays and Saturdays. All participants will be enrolledafter written, informed consent.
**Paediatrics:**Allasymptomatic paediatric participants from who are CHIKV IgG antibody positivewithout clinical manifestation will be enrolled as asymptomatic cases.Symptomatic confirmed paediatric cases (NVBDCP) will be enrolled from theDepartment of Paediatric. Age and sex matched paediatric controls will be takenfrom those relatives who accompany patients to the Therapeutic Drug MonitoringOPD run by the Department on Fridays and Saturdays. All participants will beenrolled after written, assent and informed consent from the LAR.
*Study procedure***:** A total of5 mlof blood will be collected from all study participants for assessing the CHIKVantibody. This will only be a qualitative estimation [yes/no]. There will betwo study visits (Visit 1 - [Screening] + visit-2 [collection of blood for proteomicanalysis]). If a participant who is asymptomatic from a potential control group[since at the point of screening we will not know whether a “control†willtests positive or negative for the antibody] and tests positive (>11standard units), he/she will be enrolled as an asymptomatic case andparticipants who fulfil the confirmed case definition (NVBDCP) will be enrolledas symptomatic cases. A total of 5 ml ofblood will be collected for proteomic analysis will be processed in centrifugeat 3000 rpm and plasma will be separated. Plasma will be stored under-20 degreecelsius till shipment to central laboratory (IIT Bombay). Details of theproteomics analysis are given in Appendix 1.
**Statistical Analysis:**
Descriptivestatistics (mean and SD, median and range and percentages) will be used topresent the quantitative and qualitative data. Odds ratio will be used as ameasure of association between the proteins seen in cases versus controls. Chisquare or Fisher exact test will be used for categorical data to establish the strengthof the association. A p value of less than 5% will be considered as statisticallysignificant. Graph Pad statistical software version 3 will be used forstatistical analysis.
**Anticipated outcomes:**
Basedon the previous studies and Plasma Proteome Database, at least 500 proteins areexpected to be identified with minimum of 50 proteins that will be differentiallyexpressed (over-expression and under-expression). These differentiallyexpressed protein may help to pinpoint potential biomarkers that could in thelong-term lead to development of newer vaccines that are driven by thesebiomarkers.**Background and Rationale:**Chikungunyais a viral disease transmitted by Aedes mosquitoes. Chikungunya virus (CHIKV)is an arbovirus that belongs to the family of Togaviridae and genus Alphavirus.The virus was first discovered in 1952 and used tocause sporadic outbreaks.
Inrecent years, there have been several reports of large scale outbreaks ofChikungunya virus infection in several parts of Southern India.Over,2000 cases of Chikungunya fever have also been reported from Malegaon town inNasik district, Maharashtra state, India between February-March 2006.In October 2009, 2–3 months after an outbreak of a febriledisease with joint pain on the eastern coast of Madagascar, serologic markersfor chikungunya virus (CHIKV) in n= 1,244 pregnant women were assessed and IgGsero-prevalence against CHIKV was 45%. The recent outbreaks of this diseasehave been associated with fatalities and chronic and persistent disabilityparticularly the persistence of joint pain. Chikungunya is thus considered asan important re-emerging public health problem in both tropicaland temperate countries, where the distribution of the Aedes mosquito vectorscontinues to expand.
Giventhis backdrop, several pharmaceutical companies are in the process ofdeveloping vaccines to combat the disease. One such vaccine has been developedby Bharat Biotech and is undergoing clinical development at the KEM Hospital. APhase I study [a study in normal, healthy participants is currently underway atthis institute after regulatory and Ethics Committee approval- EC-PHARMA-10/16].
Whilescreening normal, healthy participants for inclusion into this study the studyteam found 13/53 [25%] participants to be positive for anti- Chikungunyaantibodies by an ELISA test. When questioned, these 13 participants did notgive any past history of Chikungunya infection indicating that theysuffered from asymptomatic infection.When the team did aliterature review of the prevalence of anti-Chikungunya antibodies in thegeneral population, there were no studies. Only three studies on antibodyprevalence have been done in the country and these are largely confined topatients who have been hospitalized with a diagnosis of Chikungunya.
A diseaselike Chikungunya that has seen a re-emergence would benefit from a biomarkerthat facilitates its diagnosis and helps assess prognosis. Biomarker studiesbeing with an assessment of plasma proteins that are up or down regulatedduring and after the infection – an analysis that can be done by proteomics- astudy of an entire set of proteins that are modified by an organism or system.
Thus,the present study is planned with the objective of carrying out a quantitativeproteomic analysis of patients who are positive foranti-Chikungunya antibodies including those from the above-mentioned study(EC/PHARMA-10/16) as well as those who have active infection and analyseproteins that are upregulated or downregulated in them.
**Objective:**
Toassess via a quantitative proteomic analysis the differential expression ofproteins in Chikungunya antibody positive patients (symptomatic andasymptomatic) versusage and sex matched healthycontrols.
**Methods:**
*Ethics*:The study will be submitted to the Institutional EthicsCommittee for approval. Written, informed consent will be taken from allparticipants. A copy of the submissionform will be submitted to the Research Society
*Study design*:Cross sectional
SampleSize calculation: No formal sample size calculation
**Adults:**The department is currently doing a studyEC/PHARMA-10/2016 where a new Chikungunya vaccine is being tested in normal,healthy participants. A total of n = 25 participants who are asymptomatic buttested positive (>11 standard units), n = 25 who satisfy the confirmed casedefinition of Chikungunya (NVBDCP) and n = 50 controls who have tested negative(<11 standard units) will be enrolled in this cross-sectional study.
**Paediatrics:**A total of n = 10 participants who are asymptomatic but tested positive (>11standard units), n = 10 who satisfy the confirmed case definition ofChikungunya (NVBDCP) and n = 20 controls who have tested negative (<11standard units) will be enrolled in this cross-sectional study.
*Screening*:All participants will be screened (via medical history and physical examinationonly) to establish the eligibility criteria as outlined below. This includesnegative antibodies, CHIKV IgG, by ELISA method.
*Selection criteria***:**
**Adults:**Allasymptomatic participants from the Chikungunya vaccine trial (EC/PHARMA-10/16),aged ≥18 and ≤50 years and who are anti-CHIKV antibody positive (>11standard units) will be enrolled as asymptomatic adult cases. These are theparticipants who have been excluded from the vaccine study due to antibody positivity.Symptomatic confirmed adult cases (NVBDCP) will be enrolled from the Departmentof Medicine. Age and sex matched adult controls will be taken from thoserelatives who accompany patients to the Therapeutic Drug Monitoring OPD run bythe Department on Fridays and Saturdays. All participants will be enrolledafter written, informed consent.
**Paediatrics:**Allasymptomatic paediatric participants from who are CHIKV IgG antibody positivewithout clinical manifestation will be enrolled as asymptomatic cases.Symptomatic confirmed paediatric cases (NVBDCP) will be enrolled from theDepartment of Paediatric. Age and sex matched paediatric controls will be takenfrom those relatives who accompany patients to the Therapeutic Drug MonitoringOPD run by the Department on Fridays and Saturdays. All participants will beenrolled after written, assent and informed consent from the LAR.
*Study procedure***:** A total of5 mlof blood will be collected from all study participants for assessing the CHIKVantibody. This will only be a qualitative estimation [yes/no]. There will betwo study visits (Visit 1 - [Screening] + visit-2 [collection of blood for proteomicanalysis]). If a participant who is asymptomatic from a potential control group[since at the point of screening we will not know whether a “control†willtests positive or negative for the antibody] and tests positive (>11standard units), he/she will be enrolled as an asymptomatic case andparticipants who fulfil the confirmed case definition (NVBDCP) will be enrolledas symptomatic cases. A total of 5 ml ofblood will be collected for proteomic analysis will be processed in centrifugeat 3000 rpm and plasma will be separated. Plasma will be stored under-20 degreecelsius till shipment to central laboratory (IIT Bombay). Details of theproteomics analysis are given in Appendix 1.
**Statistical Analysis:**
Descriptivestatistics (mean and SD, median and range and percentages) will be used topresent the quantitative and qualitative data. Odds ratio will be used as ameasure of association between the proteins seen in cases versus controls. Chisquare or Fisher exact test will be used for categorical data to establish the strengthof the association. A p value of less than 5% will be considered as statisticallysignificant. Graph Pad statistical software version 3 will be used forstatistical analysis.
**Anticipated outcomes:**
Basedon the previous studies and Plasma Proteome Database, at least 500 proteins areexpected to be identified with minimum of 50 proteins that will be differentiallyexpressed (over-expression and under-expression). These differentiallyexpressed protein may help to pinpoint potential biomarkers that could in thelong-term lead to development of newer vaccines that are driven by thesebiomarkers.
- Detailed Description
Not available
Recruitment & Eligibility
- Status
- Not Yet Recruiting
- Sex
- All
- Target Recruitment
- 140
Able and willing to complete informed consent process with understanding of the purpose and procedures of the study Medical history and physical examination without clinically significant findings at the time of screening.
Pregnant and lactating women History of active viral infection History of immune deficiency or autoimmune disease Have active or history of arthritis within the last 6 months from date of screening Have an active or history of malignant conditions including haematological malignancy Have a history of degenerative neurological disease Have received any vaccination within 4 weeks prior to this study Participants from the vaccine study who have received the Chikungunya vaccine Have received blood transfusion or immunoglobulin within 30 days prior to vaccination in this study.
Study & Design
- Study Type
- Observational
- Study Design
- Not specified
- Primary Outcome Measures
Name Time Method Based on the Plasma Proteome Database the end point of the present study is to identify at least 500 proteins among which 50 proteins are expected to be differentially expressed These differentially expressed proteins may help to discover newer vaccine for chikungunya infection in near future Proteomic analysis of plasma protein will also identify surrogate marker for diagnosis and prognosis September 2019
- Secondary Outcome Measures
Name Time Method Based on the Plasma Proteome Database the end point of the present study is to identify at least 500 proteins among which 50 proteins are expected to be differentially expressed These differentially expressed proteins may help to discover newer vaccine for chikungunya infection in near future Proteomic analysis of plasma protein will also identify surrogate marker for diagnosis and prognosis 12 months
Trial Locations
- Locations (1)
Seth GSMC and KEM Hospital
🇮🇳Mumbai, MAHARASHTRA, India
Seth GSMC and KEM Hospital🇮🇳Mumbai, MAHARASHTRA, IndiaNithya GogtayPrincipal investigator02224133767njgogtay@hotmail.com