S-12-2 Is there a role for neurosteroids in alcoholism?

5-12 New htologtcal and pharmacotherapeutic approaches in alcoholism

192 to the illness can be rulec~ o ~ t on the bas,s of tne ong known ,,-'ned ate (but transient) amelioration induced oy sleep deprivation Instead, the delayed onset of actton appears to be related to the adaptive phenomena underlying the antidepressant response. One such important phenomenon that has been particularly well documented is the desensitization of soma~ todendritic 5-HT1A adtoreceptors during the course of an SSRI treatment which allows 5 HT neurons to recover a normal firing actwlty. The search for antidepressant agents endowed with more raptd onset of action has led to encompass the noradrenerglc(NA) system indeed.~tl/as long been known that the 5-HT and NA systems are intimately ,ntercon nected In particular. 5 - H T n e u r o n s o f d o r s a l r a p h e r e c e w e a t o r r l c e x c t a t o r y input from the locus coeruleus Venlafaxine is a dual 5-HT/NA reuptake irrhioltor w~th a preferential eIfem (in the order of 1:4) fo~ the 5 H T cart,dr Some studies have reported a rap~d o r s e t of action n patients #orwhom a rap~d titration was achieved. Gven the lower affinity of ven afax~ne for the NA carrier, ~t can be specuiateo that the recruitment of the NE system occurs only when doses in the hlgne, range ere attained Hence, ~t is possible that the above menhoned early onset of action involves the early recruitment of :he NE system This m~ght indeed results ~n synergistic effects on the 5-HT and NA systems and/or r a earher acbvat~on of the 5-H~ system through NE reuptake blockade Botr fundamental and clinical studies are underway at our research s~te to better define and understand this ootenbal y important propr ety of t~e dual B HT/NE reuptake inhibitor venlafaxlne

S-12 New biological and pharmacotherapeutic approaches in alcoholism I S°1

2-1 i Animal models in alcohol research

syndrome Tnus sP rats appear to be able to avoid the ingestion ofintoxicat'ng doses of ethanol, possibly adjusting their ethanol consumption not to exceed their rate of alcohol metabolism Over the last decade, some genetically-controlled neurochemical and oeqaworal traits, likely associated to the development of ethanol preference and dependence, have been identified in sP and sNP rats. For instance, the metabolism of dopamine, a candidate neurotransmitter m the mediation of the reinforcing properbes of ethanol, in discrete brain areas has been found to be more sensitive to ethanol-induced stimulation m s P t h a n i n s N P r a t s Moreover, lower density of t h e d o p a m i n e D 1 and D 2 receptors has been assessed in sP than in sNP rats. More recently, sP and sNP rats have been shown to differ in their anxiety profile In :his study, the spontaneous explorationoftheopen(hostilespace) and closed (comfortable space) arms of an elevated plus maze was used as a behavioral measure of anxiety Time spent in and number of entries into the open arms were lower Ln ethanol-naive sP than in ethanol-naive sNP rats, indicating inherent higher levels of anxiety in sP than in sNP rats and strengthening the hypothesis that anxiety may constitute a critical factor in the ebology of ethanol preference However, voluntary ethanol intake for 14 consecutive days partially reversed the anxiety profile of sP rats. Indeed, t~me spent in and number of entnes into the open arms were higher in ethanoPexperienced than in ethanol-naive sNP rats. This result suggests that the anxiolytic effects of ethanol are part of its reinforcing properties and sP rats tend to self-medicate anxiety by ethanol drinking. F~nahy, it has been shown that sP rats possess high predictive validity the screening of drugs for their efficacy in the pharmacotherapy of al::ohoi dependerce For instance, sP rats have been successfully used in Lhe idenhfication of the promising therapeutic agent, V hydroxybutyrie acid ,GHB)

~--12-~

Is there a role for neurosteroids in alcoholism?

E Romeo, E. Pompilil, F. d i M i c h e l e ! , C r-urnaril, R F u c c i l , A Pasinil. G i a n c a r l o C o l o m b o l , q o b e r t a A g a b o 2 Nadejda 3alaklievskala2 Carla L o b i n a l Roberta Reali2 Fabo Fadda3 Gian L u g ' G e s s a 2 " C,\'R

Center for Neuropharmaco/ogy, Cagtiarr Italy, 2 "Bernard B Brod~e" Department of Aleurosc~ence, Umvers~P/ of Cag/~arr: /tatT, 3 !nst~tute ~:f Human Physiology,, University of Cagl,'arL Italy It is weil know!l that laooratory a ",lnais can be selecUve y bred to- J verger< ethanol preference and consumption Over :']e iast thirty years f,ve pal's of rat lines have been successfully selected for their difference in voluntary e t h a n o intake Smcethese~ector" D r e s e d u r e h a s b e e r ] d e s g r e d l r ] o ' d e ' t o obtain a differential segregation only at those genes involved ,n the regulat, on of ethano prefe-ence these ethanol pre;ernng ard -non p'efernng an reals have yielded coqsideraole clarification of the genetiea ly t'ansmltted determinants ared sposirg to emanol preference and dependence Ethanol-prefer'ing and non prefer-ng rats co']stltute a p o w e d u E:n,r~ al model to address stades on th,: r~euroolologcal nlecnanls~s tha: re, cer ethanol dleasuraole and craved oy some ~rldlvlduals leading them to excessive consu~rption, and aversive and nox,ous Ir" others determlrl rig conqp eta ,election cf alcohohc eeverages Two lines o; ethanol preferr ng and non prefemn 3 rats have bee' se !acted n our laboratory since 1981 ]hese rat lines have beer ~arned 'Sat d nian alcohol preferr 'sq" (s p} ar/o 'Sa~dirlian a cobol-nor orefernn 9' (;NP, ressectively The se~ectlor cf :s~ a~c sNP rats started from a seteroge neous base p'opulatlon of unse ended ~'lstar rats ann has been comet1 o i l by systemat c mabng repeated generation afte" gerieratlon, of ,,he "vgnest drinking rats iP the sP line and of ',he owest ermklng rats hi the sNP h';e Ethanol preference in sP ard sNP ~s evaluatec under tee two~bott!e free choice reg men when the rats are 2 to 3"~ontbs o!d Nar-~ey rats a~( r~d viduaily housec and offered two :sort e s coqtatr'lrlg ethanol sob,t( r" ['()~; v/v) and tap water, ~espect:vely ::Odd is oval!no e ad i b sP rals co'lsu~ a daly more than 5,0 g/kg etharo~ w h l e sNP rats drnk less tha~ " 0 g~kg ethanol per day More than 90°xt o; total daily E:'-~arol :Jr,n~n:g ~'1 s p rats ~,sua~v ncc,rs durng the nocturnal phase of t~e light/dark cycle and ~s spread over three or tour separate binges Pharmacl)log callv relevant amdurts of ethano are ingested at each olnge, as ir]eica:ed oy oiood ethano concentrat ors ,~n,ut" peak uu to 80 mg°/c The daily voluntary ntaKe of at' anol m sP rats remains stable even w h e , ethanol is contlrlUOuSly offered for several months However. the pro or]gee free-choice drinking dOeS not lead to develoE)ment of physical des)andante Irdeed, abrupt remora, cf ethan{) after 12 consecutwe months u'idef the two-bottle regimen did not resJIt , any discernible sgr" o; ,.,v,thdrawal

D/pafSJmeeti d/Med/cJna Sperimentafe e Scienze Biochim/ohe e di San/to' Pubbhca, Universita" /-or Vergata, Via Tot Yergata 135, Roma 00133, Italy Several lines of evidence ~ndisate that the mammalian brain can synthesize steroids - - therefore called neurosteroias - - either from steroid precur sors of peripheral ong,n or independently of peripheral sources (Baulieu and Robe1 1990). G h a l c e i l s - - probably the most importantsteroidogenic ce~ls in the dra~n - - produce progesterone {PROG) and its metabolites 5Gnehydroprogesterone IDHP} and allopregnanolone {ALLO) (Campbell and
S-12 New bzological and l#~armacothcrapeu~t( appr(m('hes tn alcohoh~m at m/z 510, 496 and 343 were select,ve,, mor tO'dO for PROG, A[ ~O a : DHP r e s p e c t w e l y ( R o m e o et a ] 9 9 4 ) On the same days w e r e aa'- , n s tered auto and h e t e r o e v a l b a t i o n psvcho-~'etnc tests :o measure depression and anxiety: Hamilton Dedresslon q a t m g S c a e 8 e c k D e p r e s s ~ o n l n v e n t o r y , State and Trate A n x i e t y Inventory. q a m d t o n A n x i e t y Rat ng Staid, and a d sual And o g u e Scale {100 m m non s e g m e n t e d honzoqtal I,ne) for an×~ety (VASA) and oepress~on (VASD) Figure 1 s h o w s that the plasma leve s of ' \ h ~ O and D b P w e r e 50 to 60b'{, lower than those iiq control oat ents d u q n g the early w i t h d r a w a l (day i to b and returned to c o n t r o values on the late w i t h d r a w a l phase (days 15 anc~ 2 8 ) The d e c r e a s e of p asma A k L O a n t Crop c o n t e n t was i n d e p e n d e n t trot, major c h a n g e s of t h e p t u ~ t a r y - h y p o t h a i a ~ ' u s a e r e r a l a x ~ s function because it o c c u r r e d in a b s e n c e of significant c h a r g e s of plasma PROG leve:s De presslon and a n x i e t y scores d e t e r m i n e d by VASD and VASA were h a n d ' in the early w i t h d r a w a l penod, w h e n A_L© and DHP w e r e 'ow, but were r e d u c e c to a l m o s t normal values (Fig i ) ~r~ the late ~vlthdrawai phase T~e time course c h a n g e s of syrlrpto~-s ot anxiety anc depression determv~e:l by VASD and VASA, paralleled the t~me course c h a n g e s o b s e r v e d using the o t h e r rating sca,es H,gnly s~gnfl~aant ' egahve correlation existed b e t w e e n the s y m p t o m s of a n x i e t y a:~d deoresalon a n t the alasrna evels of A L t O and DHP (Fig 1) In a n o t h e r s t u d y ~t has deep reoorteo ti-at Ncehoi~c pabents have ,~ decrease in hver 5~ reductase act v ty iCroflholm ~t a i , 1994); t h e ' e f c r ~ a d o w n regu}ation of the enzymatic r n a c h n e w tnat s y n t h e s z e DH a aq::l ALLO from the p r e c u r s o r PROG couic be r e s p o n s b l e f o ' t h e reduceo eve, of plasma ALLO m alcohohc oat~ents We a r e n o w t e s t n g a,coho~c sub jects treated with i n c o m e t h a c n dunn:] w thorawal and prehmlnary resJ ts indicate that after i n d o m e t n a c i n acm~nlstratlon the p as'-~a levels of ALi (:, return to control values and are s g n l f l c a r t higher than Iri alcohohc~ un treated. T h e s e ;indings raise the poss~bd ty that photo]ace oglcal ~rlterven tionsalmedatmcreasingALLOcontentcoLIdbebeneflc@l nthetreat-~e': of the w i t h d - a w a l s y n d r o m e n a l c o h o l c palH'nts Fig.1 Plasma ALLO Negatively Correlate

o

VASA

VASb

•

to

Anxiety and Depression During Withdrlwal

o

Alcohol, c

pat one=;

•

193 , an,a" ts itype B C D), 9 8 % of the Caucasian population are p r e s e n t i n g t h e C " o r s The malor Dart of transferrin is synthesized in the liver, a smaller part 10%) ~n the C N S The t l y c o p r o t e m contams 6 % c a r b o h y d r a t e s which are hnked to asparglne acid, cons~stmg of m a n n o s e , galactose, N acetyl g l u c o s a r n m e and at t h e end e~alic a c i d C O " the abnormal form of transfernn, lacking partially the c a r b o h y d r a t e s : , a l c acid, galactose and N-acetyl glucosamm, can be d e t e r m i n e d by iso~iectr c focusing or ion e x c h a n g e c h r o m a t o g r a p h y f o l l o w e d b y radio immunoassay, a commercial k~t d e v e l o p e d by Kabi P h a r m a c i a A c c o r d i n g to Merature {Kab~ Pnarmacia) a cutoff of 20 d/I iq males and 26 U/I in f e m a l e s , ' d , c a t e s With a sensitivity of 9 0 % and a specificity of 9 8 % , a regular !cohol :'orsumpt~or ~ of more than 60 g/day, Carbonydrate Deficient Transfernn (COT] w a s first r e p o r t e d 1976 by H s h b i e r to oe abnormal in alcoholics with severe neurological s y m p t o m s . It ~ee~-s to fulf,II the criteria of a g o o d indicator for reguJar alcohol intake of more than 60 g alcohol/day, which describes alcohol abuse ( r e v i e w St±bier 79cN ) in oroer to vahdate the CDT, w e c o m p a r e d (Tab. 1), 20 f e m a l e and 56 n!a e aisorohcs, a d m i t t e d to a special ward f o r alcoholics of t h e Psychiatric ~Imve's~ty CImic of M u n i c h w(th the diagnosis of alcohol abuse and alcohol ::~epenoence (DSM III R ) w i t h 60 females and 47 controls. The c o n t r o l s w e r e i:at ents w th other psychiatric disorders s h o w i n g no signs of regular alcohol n t a k e Adddional!y, a g r o u p of alcohol abusers (12 females. 15 males) have r:een .west~gated during their stay in the hospital w i t h regard to t h e t i m e o~rse of the b ochemlcal parameters after alcohol w i t h d r a w a l . CDT w a s ::o:~:pareo with G a m m a Glutanyl Transferase (GGT), M e a n C o r p u s c u l a r Cell . ' c l J ~ e ( M C V ) Aspartate A m i n o Transferase (ASAT) and Alanine A m i n o :a,~sterase (ALAT) which are often abnormal in a l c o h o l i c s r~b

e1 N

Age !mean)

Alcohol consumption (g/day, meanl

CDT U/L

% CDT

,::,=~:

~,20: r," [57:

r*'c!

* I .~.0

mr4/:

427±15 4~ 4:1- 18 4 6 2 a 34 394±21

~ 7 2 5 $ 189 3080±383 <40 <40

265±68 364±60 145±14 120±12

88=2 116~21 47i04 41=04

GGT U/I

ASAT U/I

ALAT UII

MCV fl

'r"

76 9 ± 56 75 3 ± 32

20 8 ± 9 3 225 ± 4 4

947 ± 3 4 93.2 ± 1 9

Coatrol

,~r ~,7 I,(

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.

.

.

.

.

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,

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33 ~ ± 24 8 ±

15 4 56

1' 3 ±

! 7

7 6 ±

O 7

128±

17

92±

07

81

-I: 0 9

111±09

89.6 ±

1.4

87.3 ± 1.4

D i l l of I l l , ~ r l l l ~ A~LO ,mrlul VASA r - - 9 1

~<.OO1 AL.O re-,J, VASD r - - 8 2 p<~OOt :R,peatmd ~eambte Aplty~,s o' Variance

References Baulieu E E

n o b e l RJ l l )9[)1 '~ ' u r o s t e r o , s

'1, ,,,, h ' d l '

!~ / t o " }

-~tero d 810( he~':

Mol B~o137,395 403 Campbell J S and Karavolas H J ~]9891 The <.~.+ ~echaq ~m c" 'he "'vootqa amlc br gesterone 5c~-recuctase J Ste~o c B~ochem J2 283 28c! Cronho m ~ Borg 8 Vlestam Rams M 8jovall (199&) metd[" b : prot ie ol tero,,:Js unne ol alcoholics after withdrawal J Stero<: ~ochem Molec Blo149 101 105 Li£kow B I and Goodw n D W {! 987) Pharrnaco og
S-12-3

Carbohydrate deficient transferrin (CDT) in comparison to other biological markers in alcoholism

M Ackennell i, A Sanktjcm~r'r!se ~ & . : , ,dr ! '4
~he resu ts of our s t u d y s h o w that alcoholic pat ents, on average, have ~ " v elevated COT values in males, a s s u m m g a cutoff of 20 U/I. G G ~ ASAT, ,'<*\T a~d MCV w'ere abnormal as w e l l A c c o r d i n g to a statistical receiver E erat ng characterist c (ROD} analysis the sensitivity w a s h i g h e s t f o r CDT 8 ) % ! , f o l l o w e d ~y GGT (74%), M C V ( 7 0 % ) , A S A T ( 5 6 % ) , ALAT ( 4 0 % ) w h i c h - r e s p e r ] a s to l i t e r a b r e The specff city reacqed 9 6 % f o r C D T - ' i e ~se of the ROD curve and plotting the true positive p r o p o r t i o n versus • ~,~ ; a s e positive proportion for all possib!e cut off values, results in o u r t x i v v- a cut off value of 26 U/I for males and of 21 3 U/I f o r f e m a l e s A ' i o t q e r ~actor for u n c e r t a i n t y w h i c h has to be c o n s i d e r e d is t h e t i m e detv l e t w e e n the last alcohol intake and the blood s a m p l i n g The t i m e o f a b t e ' c e w a s < S d a y s Accerd~ng to o u r clinical lnterviews, t h e u n c e r t a i n t y e a r e d to time d e l a y however, should be n e g l e c t e d !n oJr longitudinal : ~:t,, o" hospitalized patients with blood s a m p l i n g s taken at admission and '~eek V ove ~ a p e n o d of 2 - 3 weeks, we f o u n d a significant d e c r e a s e of 3 2 8 :,: 1 9 3 r~ f e m a l e s a n t 3 6 7 to 1 8 2 in males ( T a b 2 ) The o t h e r laboratory , ~'k~rs c~ecreased as well, but w e r e not s , g n i f i c a n t w i t h t h e e x c e p t i o n of , N ' , a ; e r c e n t a g e of CDT to t r a n s f e r r i n b i d ' _aborater~ markers o ~ hospitalized abusers ID2} Geometric means with 95% :: " Jen,:e interval q ,I,de

~

Blood sampling 1

2

3

3 2 8 ( 2 2 2 485/

2 2 6 ( 1 5 3 335)

1 9 3 ( 1 3 3 28.1)

53 l i

25 5 (19 2 33 7)

18.2 (132 25

, e,~, },@q

: ( l~cl

t fi~ ,,5 : p kct ....

'

~; ' '

36 7 (25 3

1)

~, ( : o ' : ! ~ g to , j r results, C D T is a g o o d indicator for t h e regular conm l p t , o r of m o ' e than 60 g/oay alcohol The decrease d u r i n g t h e a b s t i n e n t ;=dr od characterizes CDT as a state marker w h o s e p a t h o p h y s i o l o g i c a l im; , ,:st-rE m relation to chronic alcohol intake is w o r t h to be investigated. T h e me' tee c a r b o h y d r a t e deficient g l y c o p r o t e i n s y n d r o m e (Jaeken et al, 1991 ) ne'¢~ n e u r o l o g c a l aonormalities defined in this r e a c t i o n O t h e r biological ~,~l.ers e g the ~ _ b n o r m a l m o n o a m l n e o x r d a s e ( M A O ) a n d a d e n y l a t c y c l a s e