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대한내분비학회지 : 제 21 권제 1 호 2006 원저 프라더윌리증후군에서성장호르몬의치료효과 아주대학교의과대학내분비대사내과학교실, 임상유전학교실 1 박지은 이승원 송경은 이형숙 김대중 정윤석 이관우 김현주 1 Growth Hormone Treatment in Prader-Willi Syndrome Ji Eun Park, Seung Won Lee, Kyoung Eun Song, Hyoung Suk Lee, Dae Jung Kim, Yoon-Sok Chung, Kwan Woo Lee, Hyon Joo Kim 1 Department of Endocrinology and Metabolism, Department of Clinical Genetics 1, Ajou University School of Medicine, Suwon, Korea ABSTRACT Background: Prader-Willi syndrome (PWS) is a congenital disorder, which is clinically characterized by a short stature, muscular hypotonia, hypogonadism, mental retardation and hyperphagia, leading to early childhood obesity. Impaired growth hormone (GH) secretion, hypogonadism, and obesity are common in patients with PWS. The purpose of this study was to find the effects of growth hormone treatment in patients with PWS. Methods: Six patients with PWS confirmed by a genetic study were recruited, and treated with growth hormone (Eutropin R ) (0.8-1 IU/kg/week) divided into five or seven day doses per week for six months. The heights and weights of the subjects were evaluated. GH status were evaluated using the serum insulin-like growth factor (IGF)-I level, the L-dopa test, and insulin-induced hypoglycemia tess. Glucose metabolism was evaluated using the random serum glucose and HbA1c levels. Results: GH was found to be deficient in 2 out of 6 subjects by the insulin test, in 3 out of 6 by the IGF-I level, and in 5 out of in 5 by the L-dopa test. After six months of GH treatment, the height percentile was increased and weight percentile decreased. The serum glucose and HbA1c levels remained unchanged. Conclusion: Six months of GH treatment in patients with PWS improved the height and degree of obesity. This study has shown the beneficial effects of GH treatment for patients with PWS, and without significant side effects (J Kor Soc Endocrinol 21:40~46, 2006). ꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏ Key Words: Growth hormone, Height, Weight, Prader-Willi syndrome 4) 서 론 프라더윌리증후군 (Prader-Willi syndrome) 은 Prader 등 [1] 에의해처음보고되었고, Zellweger 등 [2] 은 H3O증후군 (hypotonia, hypomentia, hypogonadism, obesity) 라하였으 접수일자 : 2004년 6월 11일통과일자 : 2005년 6월 16일 책임저자 : 김현주, 아주대학교의과대학임상유전학교실 * 본논문의요지는 2004년 4월 29일 ~30일대한내분비학회춘계학술대회에서발표되었음 - 40 -

- 박지은외 7인 : 프라더윌리증후군에서성장호르몬의치료효과 - 며, 현재는프라더윌리증후군또는 Prader-Labhart-Willi 증후군으로명명되고있다. 국내에서는송등 [3] 과최등 [4] 이보고한바있다. 프라더윌리증후군의정확한원인은아직밝혀져있지않았으나, fluorescence in situ hybridization (FISH), high resolution 염색체검사등의검사에서부계 15 번염색체의결손또는모계이배수체 (maternal disomy) 가관찰됨으로서 imprinting되는유전자의이상에의한유전질환으로밝혀졌다. 프라더윌리증후군환자의임상증상은발달과정에따라다르게나타나는데, 영아기에는근력저하및이에따른수유곤란과흡인성폐렴등이잘생기며, 유아기에는근력저하가호전되면서비만, 저신장, 성선기능저하, 지능저하, 비정상적인섭식습관, 작은수족등의특징적인임상양상으로나타나게된다. 이러한임상양상들중저신장, 비만, 체지방증가는성장호르몬결핍환자의임상양상과비슷하다. 실제로프라더윌리증후군에서성장호르몬의결핍이있는것으로보고된바있다 [5~12]. 프라더윌리증후군환아들에서보이는저신장, 비만, 체지방증가등의임상양상및호르몬검사상밝혀진성장호르몬의결핍으로최근문헌들에서성장호르몬치료가저신장및비만의치료에도움이됨을보고하고있다 [7~13]. 이러한보고에근거하여, 저자들은국내프라더윌리증후군환아 6명을대상으로성장호르몬결핍증여부및치료효과에대하여검토해보고자하였다. 대상및방법 2002년 8월부터 2004년 3월까지아주대학교병원유전학클리닉에서유전학검사를통해서프라더윌리증후군으로확진된환아중에서성장호르몬치료를받은 6명을대상으로후향적연구 (retrospective study) 를수행하였다. 대상자는 2.8세부터 11.1세로 6명중 4명은남아, 2명은여아이었다. 프라더윌리증후군을확진하기위한유전학적진단법으로 high resolution 염색체검사와 FISH, parental microsatellite polymorphism study를시행하였으며 methylation study로확인하였다 (Table 1). 각각의유전학적검사방법을간략히기술하면다음과같다. 임상적으로프라더윌리증후군이의심되는환자에서는먼저 high resolution 염색체검사 (400-600 band) 를시행하여염색체이상여부를검색하며, 15q의결손이의심되거나정상핵형으로나온경우에는 15번염색체의 q11-q13의미세결손을발견하기위하여이부위의 SNRPN (small nuclear ribonucleoprotein polypeptide N) gene의 specific probe을이용하여 FISH 검사를시행하였다. 다음은 15q 11-13의미세결손이어느부모에게서받은 15번염색체인지를알기위해서 microsatellite polymorphism을이용하여 15번염색체의표식자 (D15S 543,542,541,122,128,1007, 1021,165) 로 PCR하여부모의 DNA polymorphism과비교하면부계결손 (paternal deletion), 모계이배수체, 또는양친유전 (biparental inheritance) 등을확인할수있다. 또한 Table 1. Genetic Studies for Diagnosis of Prader-Willi Syndrome Deletion of 15q11-13 by FISH Parental microsatellite polymorphism study Methylation study of SNRPN gene Case 1 Negative Maternal disomy PWS methylation pattern Case 2 Positive Paternal deletion PWS methylation pattern Case 3 Positive Paternal deletion PWS methylation pattern Case 4 Negative Maternal disomy PWS methylation pattern Case 5 Positive Paternal deletion PWS methylation pattern Case 6 Positive Paternal deletion PWS methylation pattern Table 2. Serum Insulin-Like Growth Factor 1 Levels (age, sex) IGF-1 (ng/ml) Normal range Case 1 (11.1 yrs, male) 22 29-466 Case 2 (10.3 yrs, male) 78 30-289 Case 3 (2.8 yrs, male) < 8 24-152 Case 4 (7.0 yrs, male) 17 43-373 Case 5 (6.9 yrs, female) 164 43-373 Case 6 (5.1 yrs, female) 54 33-276 IGF-I, insulin-like growth factor 1. Normal range: provided by DSL kit.. - 41 -

- 대한내분비학회지 : 제 21 권제 1 호 2006 - Table 3. Growth Hormone Stimulation by L-Dopa Test Growth hormone (ng/ml) 0 min 30 min 60 min 90 min 120 min Diagnosis Case 1 < 0.5 1.88 1.78 0.76 < 0.5 Deficiency Case 2 < 0.5 2.86 1.77 < 0.5 < 0.5 Deficiency Case 3 - - - - - - Case 4 < 0.1 < 0.1 0.48 0.12 0.27 Deficiency Case 5 0.17 0.21 1.13 2.17 0.82 Deficiency Case 6 0.1 0.38 3.03 3.24 0.67 Deficiency Table 4. Growth Hormone Stimulation by Insulin-Induced Hypoglycemia Growth hormone (ng/ml) 0 min 30 min 60 min Diagnosis Case 1 < 0.5 < 0.5 0.76 Deficiency Case 2 < 0.5 2.04 4.16 Deficiency Case 3 3.05 1.39 27.76 Normal Case 4 1.27 11.87 7.05 Normal Case 5 1.94 5.41 12.72 Normal Case 6 0.85 10.83 6.09 Normal Table 5. Bone Ages in Simple X-rays Chronologic age (yrs) Sex Bone age (yrs) Case 1 11.1 Male 9.0 Case 2 10.3 Male 9.0 Case 3 2.8 Male 0.5 Case 4 7.0 Male 7.5 Case 5 6.9 Female 7.0 Case 6 5.1 Female 2.5 Fig. 1. Growth chart of boys (Case 1, 2) with growth hormone treatment. 프라더윌리증후군의원인유전자 SNRPN의 promotor영역의 CpG island의불활성화여부를확인하기위한 methylation sensitive한 PCR (MS PCR) 로검사를시행하였다. 성장호르몬분비능을검사하기위하여, 혈청인슐린양성장인자-I 농도를방사면역측정법으로측정하였다 (DSL- 2800 kit, Webster, Texas, USA). L-dopa에의한성장호르몬자극검사를시행하였다. L-dopa (Ukisen, United Pharm., Korea) 10 mg/kg을경구로투여후 0분, 30분, 60분, 90분, 120분에채혈하여방사면역측정법으로성장호르몬을측정하였다. 인슐린에의한저혈당유발로성장호르몬자극검사를시행하였다. 속효성인슐린을 0.1-0.15 U/kg을정맥주사후저혈당 (50 mg/dl 이하 ) 이되는것을확인하였고, 0분, 30분, 60분째채혈하여방사면역측정법으로성장호르몬을측정하였다. 성장호르몬 (Eutropin, LG Life Science, Korea) 을무상공급받아 0.8~1.0 U/kg/week 용량으로주 5~7회분할하여 - 42 -

- 박지은외 7인 : 프라더윌리증후군에서성장호르몬의치료효과 - Fig. 2. Growth chart of boys (Case 3, 4) with growth hormone treatment. Fig. 3. Growth chart of girls (Case 5, 6) with growth hormone treatment. Table 6. Height Velocity before and after Growth Hormone Treatment Before (cm/6months) After (cm/6months)* Case 1 1.5 4.7 Case 2 1.0 7.3 Case 3 3.5 2.5 Case 4 3.0 6.4 Case 5 3.2 6.1 Case 6 1.1 5.6 Mean 2.22 5.43 SD 1.14 1.68 *, P < 0.01 between before and after growth hormone treatment. 취침전피하주사로 6개월간투여하였다. 성장호르몬치료전후신장과체중을측정하였다. 성장호르몬치료전후무작위 (random) 정맥혈혈당및당화혈색소 (HbA1c) 를측정하였다 ( 공복으로내원하여검사하도록권유하였으나, 프라더윌리증후군환아들의음식에대한행동이상으로진정한공복이이루어질수없어무작위검사라판단하는것이올바를것같다 ). 골연령 (bone age) 을단순방사선검사로측정하였다. 결과성장호르몬분비능검사상인슐린양성장인자-I은 6명중 3 명환아에서결핍이있었고 (Table 2), L-dopa검사상은 5명중 5명에서결핍이있었으며 (Table 3), 인슐린자극검사상은 6명중 2명에서결핍이있었다 (Table 4). 프라더윌리증후군환아 6명중 4명에서골연령이지연되어있었다 (Table 5). 성장호르몬치료시작전신장은 4명의환아에서 50 백분 위수미만이었으며, 나머지 2명은 50 백분위수이상이었다. 성장호르몬치료 6개월전과치료시작시기및치료 6개월후의신장과체중의성장곡선은 Fig. 1~3과같다. 성장호르몬투여전신장성장속도가 2.22 ± 1.14 cm/6months에서 6 개월간성장호르몬투여후 5.43 ± 1.68 cm/6months로호전되었다 (Table 6). 성장호르몬치료 6개월후 6명중 5명의환아에서신장성장곡선이치료전보다상승하였으며, 체중곡선은유지되거나감소하였다. 1명은 2.8세의남아로성장호르몬치료에별반응이없었는데, 주사를투여하는부모의순응도가낮았다 ( 성장호르몬투약률이 50% 이하 ). 인슐린자극검사상성장호르몬의결핍이있었던 2명의환아 (Case 1, 2) 에서특히뚜렷한신장증가를보였다. 이후성장호르몬투여종료 6개월후측정한신장에서다시성장속도가감소함을알수있다. 당대사에영향을미칠수있는부작용을우려하여혈당과당화혈색소를측정하였다. 성장호르몬치료전과 6개월치료후의혈당은 98.3 ± 9.0 mg/dl에서 92.2 ± 8.0 mg/dl로, - 43 -

- 대한내분비학회지 : 제 21 권제 1 호 2006 - Table 7. No Evidence of Adverse Effects on Glucose Metabolism after Growth Hormone Treatment Glucose before Tx. (mg/dl) Glucose after Tx. (mg/dl) HbA1c before Tx. (%) HbA1c after Tx. (%) Case 1 88 93 5.2 5.3 Case 2 106 102 6.2 5.8 Case 3 111 79 4.7 5.4 Case 4 96 98 5.2 5.5 Case 5 99 88 5.1 6.1 Case 6 90 93 5.4 5.3 Mean 98.3 92.2 5.30 5.57 SD 9.0 8.0 0.50 0.32 No significant difference between before and after growth hormone treatment. 당화혈색소는 5.30 ± 0.50% 에서 5.57 ± 0.32% 로의미있는변화가없었다 (Table 7). 고찰프라더윌리증후군에의한저신장에대해서는대개출생후만 1세까지는정상신장을보이다가이후 50 백분위수이하가되며, 사춘기에성장분출 (growth spurt) 이없어성인이되면 5 백분위수이하가되기도한다 [14]. 비만은보통생후 6개월부터 5~6세사이에시작되며근력저하가있던시기가지난후에나타난다. 지방의분포는특징적으로주로체간, 둔부와대퇴부에나타나며손, 발에는심하지않다. 비만의주요소인은다식증으로생각된다 [2]. 이러한임상양상들중저신장, 비만, 체지방증가는성장호르몬결핍환자의임상양상과비슷하다. 또한실제로프라더윌리증후군에서성장호르몬의결핍이있는것으로보고된바있다 [5~12]. Lee 등 [15] 은성장호르몬의투여로신장과 somatomedin 농도의증가를관찰했고, 왜소증의원인으로신경분비성 (neurosecretory) 성장호르몬결핍을제기하였다. 이러한점으로미루어보아성장호르몬의결핍이전적으로저신장및비만의원인이라볼수는없지만한요인이라생각할수있다. 따라서프라더윌리증후군에서의저신장및비만의호전을초래할수있도록성장호르몬의치료가도움이될것으로생각된다. 최근의연구들에서도프라더윌리증후군환자들에게성장호르몬치료로신장성장, 체조성개선및운동능력향상을보고하고있다 [5,7~10,13,16,17]. 또한소아프라더윌리증후군에서성장호르몬의치료로키및제지방량증가, 체지방감소, 골무기질증가등이보고되었다 [18,19]. 저자들은이에유전적으로확진된국내프라더윌리증후 군환아들에게서저신장과비만의치료를위해성장호르몬치료를시작하게되었다. 성장호르몬투여후평균신장성장속도가유의하게개선되었으며, 외국의보고와유사한효과를보였다. Angulo 등 [5] 은서양인에서성장호르몬을 2년동안투여한결과신장의표준편차는치료전 -2.2SD에서 -0.8SD로호전되었음을보고하였다. Kazuo 등 [20] 은성장호르몬을 5년동안치료한결과, 37명의환아들중 34명에서신장성장속도가 5 cm/year이상으로증가하였으며, 치료전연간신장성장속도가 4.32 ± 1.74 cm에서성장호르몬치료 1년후 8.69 ± 1.91 cm로증가되었다. 일본인에서성장호르몬치료를받지않은프라더윌리증후군환아의평균최종키는남성에서 147.7 cm, 여성에서 141.2 cm이나, 성장호르몬을투여한환아들에서는남성에서 158.0 cm, 여성에서 147.7 cm으로각각 10.3 cm, 6.3 cm 정도증가되었다. 본연구에서도 6명의환아들중 5명에서신장의성장속도증가및상대적체중감소의효과를보였다. 성장호르몬치료효과가분명하지않았던환아 1명은순응도가낮은경우이었다. 또한인슐린자극검사상성장호르몬의결핍이있었던환아 2명에서성장호르몬투여후신장의성장호전이뚜렷하였으며, 성장호르몬치료 6개월후에는신장의성장곡선기울기가다시감소하는것을볼수있었다. 이를통해성장호르몬의결핍이있는경우가성장호르몬의치료에대한더좋을것으로예측할수있겠다. 본연구에서는 6개월간의치료성적을분석하였으나, 향후장기간성장호르몬투여에따른연구가필요하리라생각되며, 성장호르몬투여후골연령, 신체활동등의변화를관찰연구하는것도필요하리라생각된다. 요약연구배경 : 프라더윌리증후군은유전적질환으로안면기 - 44 -

- 박지은외 7인 : 프라더윌리증후군에서성장호르몬의치료효과 - 형, 영아기의근력저하, 다식증, 지능저하, 비만, 저신장, 성선기능저하등의임상양상을특징으로한다. 이러한임상양상중특히저신장및비만은성장호르몬결핍에서의양상과비슷한점에있으며, 실제로일부프라더윌리증후군환아에서성장호르몬의결핍이있음이보고된바있다. 외국의보고에의하면프라더윌리증후군에서성장호르몬의투여로치료효과가있음이확인된바있어, 국내환아를대상으로성장호르몬의치료효과를검증해보고자하였다. 대상및방법 : 성장호르몬분비능은혈청인슐린양성장인자-I, L-dopa자극검사, 인슐린자극검사를시행하여측정하였다. 방사선검사로골연령을측정하였다. 성장호르몬투여를 6개월간 0.8~1.0 U/kg/week를주 5~7회분할피하주사하였고, 신장과체중을측정하였다. 환자들의당대사를평가하기위해혈당과당화혈색소를측정하였다. 결과 : 성장호르몬분비능은인슐린양성장인자-I 검사상 6 명중 3명에서결핍이있었고, L-dopa자극검사상 5명중 5명에서결핍이있었으며, 인슐린자극검사상 6명중 2명에서결핍이있었다. 신장성장속도가 2.22 ± 1.14 cm/6months에서 6개월간성장호르몬투여후 5.43 ± 1.68 cm/6months로호전되었다. 성장호르몬치료후 6명중 5명에서신장성장속도의증가가있었으며, 체중곡선이유지되거나감소하였다. 인슐린자극검사상성장호르몬결핍이있었던환아에서신장의성장이나체중의감소가뚜렷하였다. 신장과체중곡선의변화가없었던 1예는순응도가낮았던경우였다. 성장호르몬의투여로인한당대사의이상은없었다. 결론 : 프라더윌리증후군환아에게 6개월간의성장호르몬치료는신장성장속도의증가및비만의호전을가져왔다. 향후프라더윌리증후군에서성장호르몬치료에대한장기간의추적연구및신체활동개선등에대한연구가필요하리라생각된다. 참고문헌 1. Prader A, Labhart A, Willi H: Ein syndrome von dipostas, kleinwuchs, kryptorchismus, und oligophrenie nach myotonieartigen zustand im neugeborenalter. Schwei Med Whchr 86:1260-1261, 1956 2. Zellweger H, Schneider HJ: Syndrome of hypotonia-hypopigmentia-hypogonadism-obesity (HHHO) or Prader-Willi syndrome. Am J Dis Child 115:588-598, 1968 3. 송영명, 신윤식, 신미자, 강성철 : Prader-Wiili 증후군 1 예. 소아과 23:78-85, 1980 4. 최원석, 김갑병, 류희수, 이순호, 김기수 : Prader-Willi 증후군 1예. 대한비뇨기과학회지 22:360-362, 1981 5. Angulo M, Castro-Magana M, Mazur B, Canas JA, Vitollo PM, Sarrantonio M: Growth hormone secretion and effects of growth hormone therapy on growth velocity and weight gain in children with Prader-Willi syndrome. J Pediatr Endocrinol Metab 9:393-400, 1996 6. Thacker MJ, Hainline B, Dennis-Feezle L, Johnson NB, Pescovitz OH: Growth failure in Prader-Willi syndrome is secondary to growth hormone deficiency. Horm Res 49:216-220, 1998 7. Hauffa BP: One-year results of growth hormone treatment of short stature in Prader-Willi syndrome. Acta Paediatr Suppl 423:63-65, 1997 8. Lindgren AC, Hagenas L, Muller J, Blichfeldt S, Rosenborg M, Brismar T, Ritzen EM: Effects of growth hormone treatment on growth and body composition in Prader-Willi syndrome: a preliminary report. Acta Paediatr Suppl 423:60-62, 1997 9. Eiholzer U, Weber R, Stutz K, Steinert H: Effect of 6months of growth hormone treatment in young children with Prader-Willi syndrome. Acta Paediatr Suppl 423:66-68, 1997 10. Davies PA, Evans S, Broomhead S, Clough H, Day JM, Laidlaw A, Barnes ND: Effect of growth hormone on height, weight, and body composition in Prader-Willi syndrome. Arch Dis Child 78:474-476, 1998 11. Lindgren AC, Hagenas L, Muller J, Blichfeldt S, Rosenborg M, Brismar T, Ritzen EM: Growth hormone treatment of children with Prader-Willi syndrome affects linear growth and body composition favourably. Acta Paediatr 87:28-31, 1998 12. Myers SE, Carrel AL, Whitman BY, Allen DB: Sustained benefit after 2years of growth hormone on body composition, fat utilization, physical strength and agility and growth in Prader-Willi syndrome. J Pediatr 137:42-49, 2000 13. Carrel AL, Myers SE, Whitman BY, Allen DB: Growth hormone improves body composition, fat utilization, physical strength and agility, and growth in Prader-Willi syndrome: A controlled study. J Pediatr 134:215-221, 1999 14. 송재원, 양세원, 문형로 : Prader-Willi 증후군에있어서의성장및내분비기능에관한임상적관찰. 대한내분비학회지 3:231-241, 1988 15. Lee PDK, Wilson DM, Rountree L: Linear growth response to exogenous growth hormone in Prader- - 45 -

- 대한내분비학회지 : 제 21 권제 1 호 2006 - Willi syndrome. Am J Med Genet 28:865-871, 1987 16. Carrel AL, Myers SE, Whitman BY, Allen DB: Sustained benefits of growth hormone on body composition, fat utilization, physical strength and agility, and growth in Prader-Willi syndrome are dose-dependent. J Pediatr Endocrinol Metab 14:1097-1105, 2001 17. Carrel AL, Allen DB: Prader-Willi syndrome: how does growth hormone affect body composition and physical function? J Pediatr Endocrinol Metab 14 (Suppl6):1445-1451, 2001 18. Burman P, Ritzen EM, Lindgren AC: Endocrine dysfunction in Prader-Willi syndrome: a review with special reference to GH. Endocr Rev 22:787-789, 2001 19. Carrel AL, Myers SE, Whitman BY, Allen BY: Benefits of long-term GH therapy in Prader-Willi syndrome: a 4 year study. J Clin Endocrinol Metab 87:3590-3597, 2002 20. Kazuo O, Satoru S, Atsunori Y, Nobuyuki M, Ryoichi S: Effects of 5 years growth hormone treatment in patients with Prader-Willi syndrome. J Pediatr Endocrinol Metab 16:155-162, 2003-46 -