ࡱ> jli'` "bjbj 0, b"  $h@   u u u   u u u u ;.7  :u 0"u I "u u ,u k " bbb$ bbb  The Peter Carlton Jones Award 2008 I am currently a postdoctoral researcher in Grant Churchills lab at the Department of Pharmacology, University of Oxford, previously having studied for my D.Phil. in the same lab. This work focussed on a second messenger (a compound that acts inside a cell to tell it how to respond to signals such as hormones) called NAADP, which is little understood. As such, I hadnt really envisaged a potential role in diseases becoming evident so soon. However, a discussion with Emyr Lloyd-Evans, who also works in the department, in Fran Platts group, regarding his work on the regulation of lysosomal Ca2+ content, led us to start some work investigating the possible role of NAADP in this process. The initial results of this work formed the basis of my submission for the Peter Carlton Jones Award. The abstract of the work is included here, and I was also delighted to be invited to present the work at the annual conference. The opportunity to attend the annual conference, also gave me the chance to discuss my research with others in the field always a useful process, which generates new ideas. In addition the chance to meet so many families, as well as being an enjoyable experience, gives an important reminder, that the research we carry out is really worthwhile. The Peter Carlton Jones Award acts to highlight the need for research into the Niemann-Pick diseases and I am very pleased to have had my research recognised in this way. Whilst the existing work looks very exciting, the initial results now need to be built upon, and developed to a stage where they are ready for publication, and will eventually inform us about possible targets and potentially lead compounds for therapies. I have submitted an application to the government funded Medical Research Council to fully investigate the role of NAADP in lysosomes in both normal cells, and disease states such as Niemann-Pick type C. I have to wait until the end of October for the results, but really hope to be successful this time: salary and research money for three years would definitely push this work in the right direction. In the meantime, I am looking forward to a holiday in the warmer climes of the Mediterranean! Dr Alexander Lewis A novel drug-like antagonist of the NAADP receptor, a Ca2+ release channel on lysosomes, produces a Niemann-Pick C disease phenotype, indicating a target for therapy. Alexander M Lewis, Emyr Lloyd-Evans, Frances Platt and Grant C Churchill. Department of Pharmacology, University of Oxford, Mansfield Road, Oxford, OX1 3QT. Niemann-Pick type C1 (NPC) is an autosomal recessive neurodegenerative lysosomal storage disorder. NPC is caused by mutations in a late endosomal/lysosomal protein (NPC1) of unknown function leading to accumulation of glycosphingolipids, sphingosine, sphingomyelin and cholesterol in these compartments. How mutations in the NPC1 protein leads to lipid storage and how lipid storage leads to neuropathology remains unclear. Previous work has shown that NPC1 null cells have a large reduction (~70-80%) in the late endosomal/lysosomal acidic Ca2+ pool, that leads to downstream endocytic transport defects and secondary lipid storage. NAADP (Nicotinic Acid Adenine Dinucleotide Phosphate) is the most potent known Ca2+-releasing second messenger. The NAADP receptor remains unknown, but work in both sea urchin and mammalian systems indicates that NAADP-induced Ca2+-release is from an acidic lysosomal store. As lysosomal Ca2+ is known to be altered in NPC, the functioning of the only Ca2+-release channel on the lysosomes is likely important in this disease. Research into the functions of NAADP has been greatly hampered by the lack of suitable chemical tools. However, using virtual screening we have developed a series of drug-like NAADP mimetics. One of these, Ned-14, is a drug-like fluorescent antagonist of the NAADP receptor. The trans isomer of Ned-14 has an IC50 of 3 nM. We used Ned-14 to confirm that the NAADP receptor is located on lysosomes, and then analysed the effect of the inhibition of the NAADP receptor on lysosomal function. Inhibition of the NAADP receptor in normal macrophages induced a phenotype remarkably similar to NPC. We observed a marked increase in lysosomal storage, as assessed using a lysotracker green fluorescence assay. Subsequent analysis of the storage material demonstrated that a wide range of products including sphingomyelin, cholesterol and glycosphingolipids were stored in the lysosomes (specific hallmarks of an NPC phenotype). Furthermore, we found that endocytic transport was defective following treatment with Ned-14, as lipid trafficking from lysosomes to the Golgi was inhibited in a similar manner to NPC cells. Thus, inhibition of the NAADP receptor produced a dysfunction in lysosomal function very similar to that found in NPC. Lysosomal Ca2+ levels were measured and found to be normal, suggesting that the lysosomal dysfunction occurred because of a reduction in Ca2+ release from the lysosome, rather than an alteration in the internal environment. This suggests that altering the degree of Ca2+-release from the lysosome via the NAADP receptor may be a useful method to improve lysosomal function. In NPC, lysosomal Ca2+-release with NAADP-AM (a membrane permeable version of NAADP, developed and synthesised in house) is reduced but not completely inhibited as with Ned-14. This raises the possibility that prolonged activation of the NAADP receptor, increasing Ca2+-release may improve aspects of NPC cellular dysfunction. To this end, the compounds selected in the virtual screen that produced Ned-14 are now being retested for agonist, rather than antagonist activity. As these compounds are already selected based on structural characteristics that make them suitable for use as drugs, the development of these compounds is substantially faster and less costly than those developed by conventional means. Thus this work raises not only a possible therapeutic target for NPC, but also a timely and economical method for its development. Lay summary: The role of calcium ions within cells has been extensively studied as it is involved in a diverse range of processes from fertilisation to cell death, and including the beating of your heart and firing of your brain cells as you read this text. We have studied a chemical called NAADP that is able to release calcium ions from lysosomes into cells. As a part of this work we developed a drug-like blocker of this process, called Ned-14. We found that Ned-14 causes storage in lysosomes that is very similar to that found in Niemann-Pick disease. We are therefore aiming to develop further drug-like activator chemicals that may lead to treatments for this disease. $ !MO 8:9; ""¸̸xnh,h4tCJH*h,h4tCJH*hwTh4tCJH*hwTh4tCJhwTh4t5CJH*hwTh4t5CJhS\ h4tOJQJ^Jh4tOJQJ^Jh'OJQJ^Jh@JOJQJ^JhS\ h@JH*OJQJ^JhS\ h@JOJQJ^JhS\ h@J5OJQJ^J'$% $a$gd4t$a$gd@J"-.+,  ""$a$gd4t$a$gd4t.:p@J|. A!"#$% <@< NormalCJaJmH sH tH DA@D Default Paragraph FontRi@R  Table Normal4 l4a (k@(No List\O\ |#Thesis Chapter Heading$a$gdi\,5CJ XOX |#Thesis Section Headingdgdi\,5NON |#Thesis subheadingdgdi\,6,$%l l  l l  l l  l l l l l l l l l l l l l l l l l l s$% - . +,00000000000000000000000000000000000000000"""8@0(  B S  ?    ' ) 6 { , P Z pu3333S\ nW4t'@J@\0@UnknownGz Times New Roman5Symbol3& z Arial qhˊنˊن 0 0!24d2QP '2#The Peter Carlton Jones Award 2008 S MathiesonMichelle ScobieOh+'0́ ( H T `lt|$The Peter Carlton Jones Award 2008 S Mathieson Normal.dotMichelle Scobie2Microsoft Office Word@F#@'7@'7G8Qt  @&" WMFC 8Xl^iQt EMFXxQ   ^iRp@"Arial3& z Arial wl 7888!8182782G8!2Tt1oUU@@1L^i formed the basis of my submission for the Peter 8!S888888227T1288T22788!88B88!&" WMFC X T UU@@LL^iCarlton Jones Award. The abstract of the work is included here, and I was H8!8827882CG8!8=88782!82787G8!2282888788!7778G82 T,  UU@@q PL^ialso delighted to be invited to present the work at the annual conference. The 828787888878818888!818887G8"2888887882778 7828=88 T UU@@ 3L^iopportunity to attend the annual conference, also g8878!8818878878878881878!88187287T  UU@@ L^ixave me the chance to 818T8881888188 T l UU@@W LL^idiscuss my research with others in the field always a useful process, which 822822T1!8288!18G8888!2888888G912882788!82822G828 Tp  UU@@ JL^igenerates new ideas. In addition the chance to meet so many families, as 7888!88288G8882878887782788278T8828T7818T8282 T 7R UU@@= ML^iwell as being an enjoyable experience, gives an important reminder, that the G88288878878818888188!8828718288S88!88!7T788!8878 T|V s UU@@ L^i\research!8288!28TtV  UU@@t DL^i we carry out is really worthwhile. The Peter Carlton Jones Award G828!!1882!881G8!8G88>78C88!H8!8728782CG8!8 T 8 UU@@# 8L^iacts to highlight the need for research into the Niemann8227878788887888!!8288!278778H8S888TT 8 UU@@ # L^iP-!T "8 UU@@ # L^itPick diseases and I C228288182878 T< B UU@@ ?L^iam very pleased to have had my research recognised in this way.8T18!188828888718888T1!8288!28!72878288882G81TTC< y UU@@C L^iP 7 TT # UU@@ L^iP 6 T`" UU@@| .L^iWhilst the existing work looks very exciting, a728881287G8"2882218!181387T  " 7 UU@@ | #L^ithe initial results now need to be 8888!818278G8888888 T( UU@@ OL^ibuilt upon, and developed to a stage where they are ready for publication, and 88788887878188888882878G88!88818!8!88818!8882888878 TwUU@@bFL^iwill eventually inform us about possible targets and potentially lead G818888188!S7288788822888!782887878781888 T{UU@@HL^icompounds for therapies. I have submitted an application to the governm28T7878828!78!88828818278T887887828878877818!8TTd{NUU@@L^iTent 88 T]UU@@HJL^ifunded Medical Research Council to fully investigate the role of NAADP in 88788S7828H8277!28H8882881828278888!877HBCHC8X &WMFCX TaUU@@BL^ilysosomes in both normal cells, and disease states such as Niemann12828T82888878!T82828788278282882182882H7T878TTaUU@@L^iP-!TlaUU@@L^iXPick C22 T86CUU@@.RL^itype C. I have to wait until the end of October for the results, but really hope 188H78188G888787887N1877!8!87!828288!7818888 TGUU@@L^i|to be successful this ti8782821821882TGUU@@4L^ime: salary and research money for three years would T8288!1888!8188!18S88818!8!88188 2G888 T4')UU@@QL^idefinitely push this work in the right direction. In the meantime, I am looking 87881882882G8!2878!788!8287888T788T87T88287 T- UU@@?L^iforward to a holiday in the warmer climes of the Mediterranean!8!G8!88888881888G8!T8!2S82788S888!!88788TT -UU@@ L^iP 5 TT#UU@@L^iP 6 TPUU@@mL^ipDr Alexander LewisH!C818888!88G2TTQUU@@QmL^iP 6 TT#UU@@L^iP 6 TT#hUU@@SL^iP 6 TTl#UU@@L^iP 6 TT#NUU@@9L^iP 6 TTR#UU@@L^iP 6 TT#4UU@@L^iP 6 TT8#UU@@L^iP 6 TT#UU@@L^iP 6 TT#UU@@xL^iP 6 TT#UU@@L^iP 6 TT#sUU@@^L^iP 6 TTw#UU@@L^iP 6 TT#YUU@@DL^iP 6 TT]#UU@@L^iP 6% 6i6^i6^66h6]h6]66g6\g6\66f6[f6[66e6Ze6Z66d6Yd6Y66c6Xc6X66b6Wb6W66a6Va6V6 6 `6U`6U 6  6 _6T_6T 6  6 ^6S^6S 6  6 ]6R]6R 6  6 \6Q\6Q 6 6[6P[6P66Z6OZ6O66Y6NY6N66X6MX6M66W6LW6L6  H.@"Arial- @2 u#GThe Peter Carlton Jones Award 2008          2 u8G @"Arial- 2 G y2 IGI am currently a postdoctoral researcher in Grant Churchills lab at the                   }2 LGDepartment of Pharmacology, University of Oxford, previously having studied                     t2 FGfor my D.Phil. in the same lab. This work focussed on a second messen                 2 Gger (a h  2 NGcompound that acts inside a cell to tell it how to respond to signals such as                     2 NGhormones) called NAADP, which is little understood. As such, I hadnt really                2 MGenvisaged a potential role in diseases becoming evident so soon. However, a                      #2 Gdiscussion with      2  GEmyr Lloyd    2 JG-U2 O1GEvans, who also works in the department, in Fran l             @"Arial---p2 %CGPlatts group, regarding his work on the regulation of lysosomal Cas                   -2 pG2+-2 %} G content,   |2 8KGled us to start some work investigating the possible role of NAADP in this a                     2 LGprocess.   2 LG  2 _G ;2 s GThe initial results of this work          U2 sX1G formed the basis of my submission for the Peter l             }2 LGCarlton Jones Award. The abstract of the work is included here, and I was                 2 PGalso delighted to be invited to present the work at the annual conference. The                       X2 3Gopportunity to attend the annual conference, also g               +2 Gave me the chance to       }2 LGdiscuss my research with others in the field always a useful process, which                           z2 JGgenerates new ideas. In addition the chance to meet so many families, as                  2 MGwell as being an enjoyable experience, gives an important reminder, that the h                      2 Gresearch   q2 DG we carry out is really worthwhile. The Peter Carlton Jones Award                  _2 8Gacts to highlight the need for research into the Niemann                2 G-)2 $GPick diseases and I      j2 "?Gam very pleased to have had my research recognised in this way.                 2 "jG  2 6G P2 I.GWhilst the existing work looks very exciting,            @2 I#Gthe initial results now need to be          2 ]OGbuilt upon, and developed to a stage where they are ready for publication, and                     t2 pFGwill eventually inform us about possible targets and potentially lead                  w2 HGcompounds for therapies. I have submitted an application to the governm                   2 Gent z2 JGfunded Medical Research Council to fully investigate the role of NAADP in                  n2 BGlysosomes in both normal cells, and disease states such as Niemann                    2 G-2 GPick r  2 RGtype C. I have to wait until the end of October for the results, but really hope                   /2 Gto be successful this ti      Y2 $4Gme: salary and research money for three years would              2 QGdefinitely push this work in the right direction. In the meantime, I am looking                     j2 ?Gforward to a holiday in the warmer climes of the Mediterranean!                  2 XG  2 G &2 GDr Alexander Lewis      2 G  2 3G  2 GG  2 ZG  2 nG  2 G  2 G  2 G  2 G  2 G  2 G  2 G  2 G  2 G  2 1G "System-GGGGFFFFFFFFFFFFEEEEEEEEEEEEDDDDDDDDDD՜.+,0  hp  University of Oxford0 ' $The Peter Carlton Jones Award 2008 Title !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`bcdefghkRoot Entry FpG.7m1TableWordDocument0,SummaryInformation( DocumentSummaryInformation8aCompObjq  FMicrosoft Office Word Document MSWordDocWord.Document.89q