«ARAVIND MEDICAL RESEARCH FOUNDATION Aravind Medical Research Foundation is recognized as Scientiﬁc and Industrial Research Organization (SIRO) by ...»
ARAVIND MEDICAL RESEARCH FOUNDATION
Aravind Medical Research Foundation is recognized as
Scientiﬁc and Industrial Research Organization (SIRO) by
the Department of Scientiﬁc and Industrial Research (DSIR)
To eliminate needless blindness by providing
evidence through research and evolving methods
to translate existing evidence and knowledge
into effective action.
IN OPHTHALMIC SCIENCESDr. G. Venkataswamy Eye Research Institute Annual Report 2012 - 2013
ARAVIND MEDICAL RESEARCH FOUNDATIONMuch has been done, but much remains to be done… we look to the future with renewed strength to continue the mission of providing quality eye care and hope that some of what we have learned will be useful to other eye care workers around the world.
Dr. G. Venkataswamy Eye Research Institute
ARAVIND MEDICAL RESEARCH FOUNDATION
BOARD OF MANAGEMENTMR. R.D. THULASIRAJ, DR. P. NAMPERUMALSAMY, DR. G. NATCHIAR, MS, DO ER. G. SRINIVASAN, BE, MS MBA MS, FAMS DR. R. KIM, DO., DNB DR. S.R. KRISHNADAS, DR.N.VENKATESH PRAJNA DR. S. ARAVIND, MS, MBA DO., DNB., FRCophth DO., DNB
RESEARCH ADVISORY COMMITTEE INSTITUTIONAL REVIEW BOARD (IRB)MEMBERS Chairman Chairman
DR. P. NAMPERUMALSAMY MS. SHOBHANA RAMACHANDHRANChairman - Emeritus Managing Director Aravind Eye Care System TVS Sri Chakra Ltd.
1, Anna Nagar, Madurai – 625 020 Madurai Member-Secretary Member-Secretary
DR.VR. MUTHUKKARUPPAN DR. LALITHA PRAJNADirector - Research Chief Microbiolgist Aravind Medical Research Foundation Aravind Eye Hospital 1, Anna Nagar, Madurai – 625 020 1, Anna Nagar, Madurai - 625 020 Members Members DR. C. MOHAN RAO PROF. C. SRINIVASAN Director UGC Emeritus Professor Centre for Cellular & Molecular Biology School of Chemistry, Madurai Kamaraj University Uppal Road, Hyderabad – 500 007 7th Street, Kalvi Nagar, Nagamalai, Madurai
DR. KUMARAVEL SOMASUNDARAMDR. L. THAYUMANAVA
(IIRUW WR XQGHUVWDQG H\H GLVHDVHV DQG DSSOLFDWLRQ RI WKH UHVHDUFK ¿QGLQJV DW WKH SDWLHQW FDUH OHYHO KDVbeen our major focus this year also. Apart from the continuation of the ongoing activities AMRF embarked on a major research program on fungal keratitis. This group has received major funding in the form of a program support from the Department of Biotechnology, Government of India, for fungal keratitis research.
This support allowed the institute to strengthen the Proteomics research area. The grant made it possible for the institute to procure a state of art Mass spectrometer, Oribitrap Velos Pro, for the study of proteomics of eye diseases. This will allow the proteomics group to do the deep analysis of expression and quantitation of proteins and their isoforms. Having this grant allowed the proteomics group to successfully attract funding from Cognizant industry also. This group, with faculty and senior investigators with Ph.D degree, will be a model for other groups to consolidate and attract major funding.
0ROHFXODU JHQHWLFV JURXS FRQWLQXHV WR H[SORUH WKH JHQHWLF RUJDQL]DWLRQ DQG GLVHDVH VSHFL¿F PDUNHUVfor various eye diseases. Apart from looking at known and new markers and mutations this group also
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PXWDWLRQV 7KH\ KDYH DOVR WULHG WR H[WHQG WKHLU ¿QGLQJV WR FOLQLFDO XVH ZLWK FORVH FROODERUDWLRQ ZLWK FOLQLFLDQVStem cell biology group is undertaking major efforts to understand the biology of buccal stem cells and also keeps in focus the application aspects. The new area of focus is role of micro RNAs in stem
FHOOV 7KLV LV DQ DFWLYH DUHD RI UHVHDUFK DQG KRSHIXOO\ VRPH QRYHO ¿QGLQJ ZLOO HPHUJH IURP WKLV DQDO\VLV
2FXODU SKDUPDFRORJ\ JURXS LV FRQWLQXLQJ LWV HIIRUWV WR EULQJ DERXW GUXJV EDVHG RQ WKH ¿QGLQJV LQdiabetic retinopathy and age related macular degeneration. The Human Organ Culture Anterior Segment System is now optimized and new results are emerging from this novel experimental system, since there is no in vivo system available for glaucoma studies.
0LFURELRORJ\ JURXS KDV GRQH VLJQL¿FDQW ZRUN LQ EDFWHULDO NHUDWLWLV SDUWLFXODUO\ LQ WKH GHWHUPLQDWLRQof virulence factors in Pseudomonas keratitis. This group is also exploring the role of micro RNAs in pathogenic process.
The new group on Bioinformatics has started its activities in the structural biology of single nucleotide variants. This group also helps in setting upvarious analysis platforms for NGS data. The analysis of
ZKROH JHQRPH VHTXHQFH GDWD RQ SDWKRJHQLF EDFWHULD E\ WKLV JURXS KDV KHOSHG LQ WKH LGHQWL¿FDWLRQ QRYHOfactors involved in pathogenesis.
Faculty of the institute also has taken up various teaching activities in the form of workshops and short terms hands on training activities.
MOLECULAR GENETICSOngoing research programmes of this group are related to commonly occurring eye diseases in Indian population like Diabetic retinopathy, Glaucoma, Cataract, Corneal dystrophies and Retinal dystrophies.
Currently gene therapy is available for Lebers Congenital Amaurosis (LCA). To implement gene therapy in Indian population, the department has been screening RPE65 genes mutations in LCA patients
ZKR FDQ EH EHQH¿WHG IURP WKLV WKHUDS\ LQ IXWXUH 7KHdepartment has recently initiated studies to explore mitochondrial involvement in Primary Open Angle Glaucoma and association of microRNA with keratoconus in collaboration with Queens University, Belfast,
8. 7KH GHSDUWPHQW KDV LGHQWL¿HG D FDQGLGDWH JHQH IRU RFXODU FRORERPD LQ FROODERUDWLRQ ZLWK &HQWHU IRUHuman Molecular Biology and Genetics, Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital, Sichuan, China. Recently it has also initiated the study to characterize the gene expression
SUR¿OH RI QRUPDO GLDEHWLF DQG GLDEHWLF UHWLQRSDWK\ KXPDQ GRQRU H\H EDOOV REWDLQHG IURP 5RWDU\ $UDYLQGInternational Eye Bank. The Department has also analysed several families with Retinoblastoma and made
VLJQL¿FDQW FRQWULEXWLRQV LQ WKH PDQDJHPHQW RI 5HWLQREODVWRPD E\ LGHQWLI\LQJ 5% PXWDWLRQV LQ QHZ ERUQVand treating the disease at an early stage. The department organized a workshop entitled “Vision Genomics” to motivate young researchers to involve in basic/ophthalmic research. Many of the study designs of the department emphasize the molecular mechanisms of various eye diseases at the gene expression level.
Molecular genetics of Leber Congenital Amaurosis in South Indian population Investigators : P. Sundaresan, Aravind Medical Research Foundation, Madurai P. Vijayalakshmi, Aravind Eye Hospital, Madurai Ph.D. scholar : Anshuman Verma Funding agency : Indian Council of Medical Research (ICMR), University Grant Commission (UGC) Fellowship Background and aim Leber congenital amaurosis (LCA) is the most severe form of visual impairment found in children. At present, more than twenty genes have been reported for LCA, and among them RPE65 is a likely candidate for gene therapy. The aim of this study is to perform a comprehensive screening of LCA genes emphasizing RPE65. In this study, three sequencing technologies were applied - (a) Direct sequencing to screen RPE65 mutations in 30 LCA probands (b) Microarray based comprehensive detection of all known LCA pathogenic variations in 25 LCA patients excluded for RPE65 mutations (c) Next generation sequencing (NGS) based targeted enrichment in a separate set of 25 LCA pooled DNA samples was carried out to screen coding regions of 56 genes, including15 known and 41 predicted LCA genes. In addition, in silico characterization
RI WKH LGHQWL¿HG PXWDWLRQV DQG UHODWHG FOLQLFDO SKHQRW\SH RI SDWLHQWV ZHUH VWXGLHGThe combined approach of RPE65 mutational screening with Asper chip analysis revealed ten different disease causing variations in 6 LCA genes from 11 LCA patients. The targeted resequencing of 56 genes uncovered total 97 variations, mostly in known LCA genes and A few predicted genes. However, most of
WKH YDULDWLRQV ZHUH LQ ÀDQNLQJ LQWURQLF UHJLRQV DQG IRXQG WR EH DV SRO\PRUSKLVPImplication of the data
7KLV LV WKH ¿UVW VWXG\ ZKLFK DSSOLHV DGYDQFHG WHFKQLTXHV IRU D FRPSUHKHQVLYH PXWDWLRQDO DQDO\VLV RILCA genes in South Indian cohort. The combined approach of direct sequencing and Asper chip analysis was instrumental to identify pathogenic mutations in 36.6 % of the patients with the major involvement of RPE65 (16.6%), GUCY2D (10%), RPGRIP1 (3.3%), AIPL1 (3.3%), CRX and IQCB1 (3.3%). In this study, RPE65 mutations were found to be the main cause for LCA followed by GUCY2D. The NGS based targeted resequencing can be productive for the comprehensive screening of a large number of genes but
DUH XQIDYRXUDEO\ LQÀXHQFHG E\ YDULRXV VHTXHQFLQJ VWHSV LQ SRROHG VDPSOHV UHVXOWLQJ LQ LWV OLPLWDWLRQ WRidentify rare pathogenic variations.
Molecular genetics and functional analysis of candidate genes associated with microphthalmia, anophthalmia and coloboma Investigators : Dr. P. Sundaresan (Aravind Medical Research Foundation, Madurai) Dr. P. Vijayalakshmi (Aravind Eye Hospital, Madurai) Dr. S. K. Kedia (Sadar Hospital, Ara, Bihar) Collaborator : Dr. Yang Zhenglin, Sichuan Academy of Medical Sciences, China PhD scholar : Mr. Sushil Kumar Dubey Funded agency : ICMR Background and aim Microphthalmia, anophthalmia and coloboma (MAC) are a group of congenital eye diseases that represent important causes of paediatric blindness. These ocular disorders have variable and poorly understood effects that extend all the way to total blindness. Extensive studies of these ocular disorders have reported mutations in a large number of genes and also several chromosomal aberrations in various populations.
Lack of similar studies for Indian population limits the knowledge regarding the genetics of these globe
DQRPDOLHV 7KLV VWXG\ DLPV WR ¿QG RXW WKH VSHFWUXP RI JHQHWLF YDULDWLRQV LQ FDQGLGDWH JHQHV DQG WKHLU UROHin causing disease.
DNA samples of 65 MAC patients were evaluated for mutations and sequence variants in the candidate
JHQHV E\ GLUHFW VHTXHQFLQJ DSSURDFK $OO WKH H[RQV DORQJ ZLWK ÀDQNLQJ H[RQ LQWURQV ERXQGDULHV RI RAX,OTX2, SOX2 and ABCB6 genes were screened in all MAC samples. Hundred ethnically-matched healthy
FRQWURO VDPSOHV ZHUH DOVR VHTXHQFHG WR FRQ¿UP WKH LGHQWL¿HG YDULDQWV DV SDWLHQW VSHFL¿F &RPSDUDWLYHsequence analysis of RAX, OTX2, SOX2 and ABCB6 genes with their homologs across different species
ZDV GRQH WR GHWHUPLQH ZKHWKHU WKH PXWDWLRQV DUH SUHVHQW LQ WKH FRQVHUYHG GRPDLQV 0XWDWLRQ LGHQWL¿HG LQ$%&% JHQH DW $05) ZDV IXQFWLRQDOO\ FKDUDFWHUL]HG LQ =HEUD¿VK E\ 6LFKXDQ 3URYLQFLDO.H\ /DERUDWRU\ IRU +XPDQ 'LVHDVH *HQH 6LFKXDQ $FDGHP\ RI 0HGLFDO 6FLHQFHV $I¿OLDWHG +RVSLWDO 6LFKXDQ &KLQD )LYH QRYHO PXWDWLRQV ZHUH LGHQWL¿HG LQ RAX, OTX2, SOX2 and ABCB6 genes in this cohort. A homozygous substitution mutation p. Arg179Trp, found in one bilateral anophthalmia patient, was LGHQWL¿HG LQ RAX gene. Three heterozygous mutations p. Pro142Fs [c. 426insC (in a proband with bilateral anophthalmia)], p. Thr186Fs [c. 558delC (in one proband with microphthalmia in one eye and anophthalmia in other)] and a compound heterozygote p. Gln104X, p. Gln106 His (in one patient with microphthalmia) ZHUH LGHQWL¿HG LQ OTX2. These three mutations in OTX2 gene cause premature termination of the coding VHTXHQFH 0XWDWLRQ S 9DO9DO ZDV LGHQWL¿HG LQ SOX2 gene in only one patient with microphthalmia and iris coloboma. Screening of ABCB6 JHQH LGHQWL¿HG D KHWHUR]\JRXV PXWDWLRQ S $OD7KU LQ H[RQ Comparative sequence analyses of RAX, OTX2, SOX2 and ABCB6 genes with their homologs showed that mutation sites are highly conserved across various species. To test the hypothesis that disruption of the normal function of ABCB6 FDQ FDXVH FRORERPD ]HEUD¿VK HPEU\RV WUHDWHG ZLWK ABCB6 morpholinos (MOs) showed coloboma phenotype and retarded development. These phenotypes can be rescued by the coinjection of WT ABCB6 mRNA but not by the coinjection of human mRNA containing Ala57Thr mutation (Figure 1).
&RPSDUHG ZLWK ]HEUD¿VK HPEU\RV WUHDWHG ZLWK VWDQGDUG FRQWURO 02V & DQG ' ]HEUD¿VK HPEU\RV treated with abcb6MO show coloboma and retarded development in (E and F). These phenotypes can be rescued by the coinjection of WT ABCB6 mRNA (G and H), but the phenotypes could not be rescued by coinjection of the human mRNA containing Ala57Thr (K and L) mutation. This is the result of the work carried out by collaborator Sichuan Academy of Medical Sciences, China Implication of the project )LYH QRYHO PXWDWLRQV LGHQWL¿HG LQ RAX, OTX2, SOX2 and ABCB6 suggest the role of these genes in ocular development. Three mutations in OTX2 gene cause premature termination of the coding sequence. These can be a pathogenic change as the protein conformation may get altered leading to loss of function. Morpholino knockdown and rescue studies of ABCB6 LQ =HEUD¿VK GHPRQVWUDWHG WKDW PXWDWLRQV LQ ABCB6 gene might cause coloboma. Further in silico and in vitro studies of the normal and mutant protein will unravel the role of the mutations in disease development.