Am. J. Hum. Genet. 51:1089-1102, 1992

X-linked Nephrogenic Diabetes Insipidus: From the Ship Hopewell to RFLP Studies Daniel G. Bichet,* Geoffrey N. Hendy,t Michele Lonergan,* Marie-Fransoise Arthus,* Sophie Ligier, * Zdenka Pausova, t Rudiger Kluge, * Hans Zingg, t Paul Saenger, * * Ellen Oppenheimer, * * David J. Hirsch, t t Simone Gilgenkrantz, t$ Jean-Pierre Salles,§§ Isabelle Oberle,##"' Jean-Louis Mandel,## Martin C. Gregory, liii T. Mary Fujiwara, Kenneth Morgan,§ and Charles R. Scriverill# Unite de Recherche Clinique, Centre de Recherche et Service de Nephrologie, D6partement de Medecine, H6pital du Sacr6-Coeur, Universit6 de Montreal; tCalcium Research Laboratory and tLaboratory of Molecular Endocrinology, Royal Victoria Hospital and McGill University; §Department of Epidemiology and Biostatistics and Department of Medicine, IlDepartment of Pediatrics, and #Department of Biology, McGill University, Montreal; * * Department of Pediatrics, Albert Einstein College of Medicine, New York; ttDepartment of Medicine, Dalhousie University, Halifax; ttService de Genetique, Universit6 de Nancy, Nancy; §§Service de Medecine Infantile, Centre Hospitalier Purpan, Toulouse; 1I1IDivision of Nephrology, Department of Medicine, The University of Utah, Salt Lake City; and ##Unite de Biologie Moleculaire et de Genie

G&n6tique, Strasbourg

Summary

Nephrogenic diabetes insipidus (NDI; designated 304800 in Mendelian Inheritance in Man) is an X-linked disorder with abnormal renal and extrarenal V2 vasopressin receptor responses. The mutant gene has been

mapped to Xq28 by analysis of RFLPs, and tight linkage between DXS52 and NDI has been reported. In 1969, Bode and Crawford proposed, under the term "the Hopewell hypothesis," that most cases in North America could be traced to descendants of Ulster Scots who arrived in Nova Scotia in 1761 on the ship Hopewell. They also suggested a link between this family and a large Mormon pedigree. DNA samples obtained from 13 independent affected families, including 42 members of the Hopewell and Mormon pedigrees, were analyzed with probes in the Xq28 region. Genealogical reconstructions were performed. Linkage between NDI and DXS304 (probe U6:2.spl), DXS305 (St35-691), DXS52 (St14-1), DXS15 (DX13), and F8C (F814) showed no recombination in 12 families, with a maximum lod score of 13.5 for DXS52. A recombinant between NDI and DXS304, DXS305, was identified in one family. The haplotype segregating with the disease in the Hopewell pedigree was not shared by other North American families. PCR analysis of the St14 VNTR allowed the distinction of two alleles that were not distinguishable by Southern analysis. Carrier status was predicted in 24 of 26 at-risk females. The Hopewell hypothesis cannot explain the origin of NDI in many of the North American families, since they have no apparent relationship with the Hopewell early settlers, either by haplotype or by genealogical analysis. We confirm the locus homogeneity of the disease by linkage analysis in ethnically diverse families. PCR analysis of the DXS52 VNTR in NDI families is very useful for carrier testing and presymptomatic diagnosis, which can prevent the first manifestations of dehydration. Introduction

Nephrogenic diabetes insipidus (NDI; designated 304800 in Mendelian Inheritance in Man [McKusick Received April 2, 1992; revision received July 6, 1992. Address for correspondence and reprints: Daniel G. Bichet, M.D., Research Centre, Hopital du Sacre-Coeur de Montreal, 5400, boulevard Gouin ouest, Montreal, Quebec, Canada H4J

lCS.

1. Deceased, November 17, 1991.

by The American Society of Human Genetics. All rights reserved. 0002-9297/92/5105-0019$02.00 C 1992

1990]) is an X-linked disorder in which the affected patients do not concentrate their urine after the administration of the antidiuretic hormone arginine-vasopressin (AVP) (Reeves and Andreoli 1989). In this disease, the vasopressin signal is not recognized - or its transmission is altered - by the principal cells of the renal collecting ducts. The defect is not confined to the kidneys, since affected male patients have no vasodepressor or factor VIII stimulation response to

the ad-

ministration of the antidiuretic vasopressin agonist

1089

1090

1-desamino[8-D-arginine]vasopressin (dDAVP) (Kobrinsky et al. 1985; Bichet et al. 1988, 1989, 1991). By linkage analysis, the NDI gene has been assigned to the Xq28 region (Kambouris et al. 1988; Knoers et al. 1988a, 1988b, 1989). Genetic linkage studies have been carried out in one North American family and in several European families. The odds of linkage between Xq28 marker DXSS2 and the NDI locus are greater than 1013:1 (Kambouris 1988; Knoers et al. 1988b). The availability of Xq28 markers provided us with the opportunity to test a hypothesis put forward by Bode and Crawford (1969). They proposed that NDI patients in eastern North America shared a common ancestor who was an Ulster Scot and who had arrived in Halifax in 1761 on the ship Hopewell. A link between this family and a large Mormon pedigree (Cannon 1955) was also suggested. Our first objective was to test the assumption that the chromosomal location of the altered gene responsible for the disorder was identical in all affected families-i.e., locus homogeneity. A total of 13 families were studied: representatives of the Hopewell pedigree (Bode and Crawford 1969), the Mormon pedigree (Cannon 1955), and 11 unrelated families from diverse ethnic backgrounds (5 French-Canadian, 1 African-American, 1 Puerto Rican, 1 Iranian, 2 French, and 1 English). The phenotypic characteristics of affected persons were similar in all families. Our second objective was to test the Hopewell hypothesis by using DNA probes: if recombination in the Xq28 region is rare and if the mutant NDI alleles in the Hopewell pedigree and other North American families are identical by descent from an Ulster Scot ancestor, then the haplotypes of the Xq28 loci segregating with the disease should be identical in these families. Our third objective was to determine whether the heterozygotes in our pedigrees could be identified by DNA analysis, since carrier detection by other means has been difficult up to now (Bode and Crawford 1969; Bichet et al. 1988, 1989).

Subjects and Methods Study Subjects Thirteen families with well-documented X-linked NDI who were distributed in distinctive geographical, historical, and ethnic groups were studied. All affected male patients had a documented lifelong history of polyuria and polydipsia, normal or elevated plasma

Bichet et al.

concentrations of arginine-vasopressin, and unconcentrated urine despite administration of argininevasopressin or dDAVP. Phenotypic characteristics were similar within and between families (Bichet et al. 1988, 1989, 1991). Mental retardation correlated positively with delay in diagnosis and with the severity of clinical course. The phenotypic characteristics of 14 male patients and 12 obligate carriers have been described elsewhere (Bichet et al. 1988, 1989, 1991). We studied 38 affected males, 18 nonaffected male relatives, 30 obligate carriers, and 26 females at risk for heterozygosity. The families studied are shown in figures 1-4. The Hopewell pedigree (fig. 1) was reconstructed to a depth of six generations, from detailed bibliographical records (Eaton 1912; Miller 1972), from information given by family members, and from the Nova Scotian Archives (Halifax). DNA was obtained from three related families (A-C) of the Hopewell pedigree (fig. 1). Families A, B. and C were depicted, respectively, in the central, paracentral right, and right parts of the genealogy originally presented by Bode and Crawford (1969). Historical records were reviewed, and blood samples obtained from the Mormon family (Cannon 1955) (fig. 2). The five French-Canadian families (Quebec [Ql-Qs]; fig. 3) had been established in Quebec since the 18th century; their genealogies were reconstructed from Quebec archives. The other families (other [01-06]; fig. 4) reside in Quebec, Ontario, New York, and France. Family 01 is Iranian in origin, 02 is Puerto Rican, 03 and Os are French, 04 is British, and 06 is African-American. DNA, PCR, and Linkage Analyses DNA was extracted and purified from blood samples by a conventional phenol-chloroform technique (Sambrook et al. 1989) and was cut with different

restriction enzymes (New England Biolabs, Beverly, MA) in the presence of 2 mM spermidine. After electrophoresis (0.9% agarose, TAE buffer) (Sigma, St. Louis) Southern blots were performed (0.45 gm; Nytran, Schleicher and Schuell, Keene, NH) with 10 x sodium chloride-sodium citrate (pH 7.0). The DNA probes were labeled with [a-32P]dCTP (ICN Radiochemicals, Irvine, CA) by using the random primer method (Multiprime DNA labeling systems; Amersham, Oakville, Ontario). The filters were

autoradiographed (X-Omat AR5, Kodak) at - 80°C

with 2 intensifying screens (Lightning Plus; DuPont) for 4-7 d. The DNA probes (Harper et al. 1984; Oberle et al.

Insert

A

Insert

B

VII 0, IX

2

52 53 2

l

22

5

54 12

5 1 2

2

21

6 23 45 see PCR analysis fig 7

ixViIIViIVI V IV 1II IV

VIVIIVIIIIx

v

PROBES

LOCI

St35- 691 Stl4- 1 DX13 F814

VA'S305

5 1 2

718 9

/2XS52 /2XS15 FIIC

Insert C

11

11

45 22 11

55 12 21

_ -

5

2

11

1

55 12 21

5 1 2

Pedigree of the Hopewell family, segregating for X-linked NDI and Xq28 RFLP markers. Generations are numbered Figure I according to the protocol of Bode and Crawford (1969). The couple at the origin of the inserts is 111-9 and III-10 (also see fig. 5). Probes and loci are listed from centromere to telomere. In females in whom the inheritance of paternal and maternal alleles can be ascertained, the paternal X chromosome is shown on the right. In this family, the disease is segregating with the 1-5-1-2 haplotype. Allele 5, marked with an asterisk (*) was subsequently identified as allele 6, by PCR. C = Unaffected male; * = affected male; ( = obligate carrier female; and = predicted carrier female. Double lines denote consanguineous marriage.

1092

Bichet et al.

1 1 4 2 2

2 2 3 1 1

1

11

42 21 21

42 2 1 21

l-1 1 1 4 2 2

3

1 1 3

1 1

1 1

1 2 33 1 1 l i

2 2 3

2 2 3

3

1 1

1 1

1 1

1

--or-12 21 34 34 12 12 11

11

PROBES

LOC/

U6:2.spl St35-691 Stl4- 1 DX13 F814

121S304 JA'S3605 DXS52

VlSi 5 F8C

Historical reconstitution of Cannon's Mormon pedigree. The disease allele could not originate from Fullmer (born in 1803). Figure 2 The disease is segregating with the 2-3-1-1 haplotype, and a recombination was observed in the male twins, between the DXS30S and the NDI loci.0) = Carrier status unknown. All other symbols are as in fig. 1.

1985; Mandel et al. 1986; Heilig et al. 1988; Vincent et al. 1989; Rousseau et al. 1990) used here are described in table 1 and are located, from centromere to telomere, according to the results of Rousseau et al. 1990. Alleles were numbered according to the protocol of the 10th International Workshop on Human Gene Mapping (Kidd et al. 1989) and, for U6:2.spl, according to the protocol of Rousseau et al. (1990). A PCR analysis of the Stl4 (DXSS2) VNTR was done by a modification of the technique described by Richards et al. (1991). We used a thermocycler and reagents from Perkin Elmer Cetus (Montreal). Taq

polymerase (2.5 U/sample; BRL, Gaithersburg, MD)

was added to a protease-inactivated mixture (DNA, nucleotide, buffer, and primers). We programmed 25 cycles (1 min at 94°C and 4 min at 60°C). Amplified products were examined on agarose gel electrophoresis: they include VNTRs of 60 bp each and a 650-bp flanking sequence (Richards et al. 1991). PCR analysis was found to be the method of choice for analyzing the Stl4 VNTR, because this method enabled the differentiation of alleles previously not distinguishable by Southern analysis (table 1). Alleles 5 (1,630 bp) and 6 (1,570 bp) are difficult to distinguish on the

Southern blots and are arbitrarily noted on figures 1-4 as allele 5, except for family O3 (allele 6 verified).

2 4 2 1

22 4 2 22

2 1 4 5 2 2 12

11

Fi

1 2 32

15 12

1 1

11 91

Q)

2 5

9

124 d± r~ ~~~1

25 n 12 22

11

24 1 2

5 2

2

2 1

2 1

1

I

1

2 2

2 5 11

2 5 12

3 1

3 1

3 1

Li

I

11

3d

12

8 1

_

(

3 1

11 58 2 1

0 ~~~~~~~~~~~~~~~~~~~~~~~12 I I1

2 8

2 1 8 5'

2 1 i2

Q3

2

2 1 8

1 2

2

2

1

5, L

2

2 8 1 2

1 4 2 1

2 8 1 2

1 1

1 1

5*4 1 2 1 1

5*4 1 2 1 1

2 22 4 8 1 1 22

Q4

I~~

Q 4

2 1

1

12

25 2

2

2 5 2

,~~~~~~ 2

1

12

* 1

4

2

2

5

PROBES

Goc/

S -35 691 St 14 - 1

12AS52

DX I'3

PIJSIS

1XS305

F8C F814 Five Quebec families with X-linked NDI. In family Q4, crosses indicate that the two affected males died early in infancy. Figure 3 All other symbols are as in fig. 1.

Bichet et al.

1094

04

02

01

2

73

1 2

03

86 21

lIl

38 21

5

_

4 4 2 2

5

1 1

2

3 2 2

7

1

1

3 2

1

1

1

8 1 1

4 2

1 1

3 2

3 2

2 4 2

22 44 22

1

1

l

1 1

1

1

C

-EEl-o ov-" |8 6 8 6 2-l 2-,/l\

8

66

2-

1l1

2:lO

86 O21

6

23l

l

2

2

2

2

7 5*

L

1 2

*

2 5* 2

05

6

4 1 6

8

8

1

2

2

l 2

5*

2

06

PROBES

LOCI

St35- 691

Stl4-

12AS305 12XS52

DX13 F814

TI/C

12XS15

Other families. In family 03, chorionic villus sampling established that the unborn child indicated by a triangle (A) was Figure 4 risk of inheriting the disease. This prediction was confirmed by the birth of a normal infant. In 06, allele 5, marked with an asterisk (*), was identified as allele 6, by PCR. All other symbols are as in figs. 1 and 2.

not at

PCR analysis was used, a posteriori, to reclassify alleles 5 and 6 (denoted by "5k," in the figures). Linkage analysis was carried out by the likelihood method (Ott 1991) using the LINKAGE programs (Lathrop et al. 1984, 1985). Dr. Mark Lathrop provided the LINKAGE programs (version 5.1a). Pub-

lished allele frequencies for the RFLPs were used (see table 1). Linkage equilibrium was assumed. The mutant allele frequency for X-linked NDI was set at .03 for the Hopewell pedigree and at 7.4 x 10-6 for the other families (see below), and a mutation rate of 10-6 was assumed.

1095

X-linked Nephrogenic Diabetes Insipidus Table I RFLPs Used in the Linkage Study SOUTHERN ANALYSIS Locus

DXS304 DXS30S

DXS52

DXS1S

...

...

....

....

PROBE

ENZYME

U6:2.spl

BanI

St35-691

TaqI

Stl4-1

TaqI

DX13

F8C . .F814 a

BglII BclI

ALLELE

Size

Frequency

(kb)

(%)

References

76 24 65 35 3,000 2,900 2,400 1,690

1,630

20

1,570 1,390 1,300 1,220

10 15