Exploring the role of the <i>WNT5A</i> rs566926 polymorphism and its interactions in non-syndromic orofacial cleft: a multicenter study in Brazil

LARA, Lorraynne dos Santos; COLETTA, Ricardo D.; Assis MACHADO, Renato; Querino Rocha de OLIVEIRA, Lilianny; MARTELLI JÚNIOR, Hercílio; de Almeida REIS, Silvia Regina; SCARIOT, Rafaela; Evaristo Ricci VOLPATO, Luiz

doi:10.1590/1678-7757-2023-0353

person Lorraynne dos Santos LARA

Conceptualization (Equal) · Investigation (Equal) · Methodology (Equal) · Validation (Equal) · Visualization (Equal) · Writing – original draft (Equal) · Writing – review & editing (Equal)

schoolUniversidade de Cuiabá, Programa de Pós-Graduação em Ciências Odontológicas Integradas, Faculdade de Odontologia de Cuiabá, Cuiabá, MT, Brasil.

0000-0002-5082-4185

person Ricardo D. COLETTA

Conceptualization (Equal) · Data curation (Equal) · Formal analysis (Equal) · Funding acquisition (Equal) · Investigation (Equal) · Methodology (Equal) · Project administration (Equal) · Resources (Equal) · Supervision (Equal) · Validation (Equal) · Visualization (Equal) · Writing – review & editing (Equal)

schoolUniversidade Estadual de Campinas, Departamento de Diagnóstico Oral e Programa de Pós-Graduação em Biologia Buco-Dental, Faculdade de Odontologia de Piracicaba, Piracicaba, SP, Brasil.

0000-0001-5285-3046

person Renato Assis MACHADO

Conceptualization (Equal) · Data curation (Equal) · Investigation (Equal) · Methodology (Equal) · Software (Equal) · Validation (Equal) · Visualization (Equal) · Writing – review & editing (Equal)

schoolUniversidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba, Programa de Pós-Graduação em Biologia Buco-Dental, Piracicaba, SP, Brasil.

0000-0002-1697-3662

person Lilianny Querino Rocha de OLIVEIRA

Conceptualization (Equal) · Investigation (Equal) · Methodology (Equal) · Validation (Equal) · Visualization (Equal) · Writing – review & editing (Equal)

schoolUniversidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba, Programa de Pós-Graduação em Biologia Buco-Dental, Piracicaba, SP, Brasil.

0000-0003-4842-5756

person Hercílio MARTELLI JÚNIOR

Conceptualization (Equal) · Methodology (Equal) · Validation (Equal) · Visualization (Equal) · Writing – review & editing (Equal)

schoolUniversidade Estadual de Montes Claros, Departamento de Odontologia, Clínica de Estomatologia, Montes Claros, MG, Brasil, e

0000-0001-9691-2802

person Silvia Regina de Almeida REIS

Conceptualization (Equal) · Methodology (Equal) · Validation (Equal) · Visualization (Equal) · Writing – review & editing (Equal)

schoolEscola Bahiana de Medicina e Saúde Pública, Departamento de Ciências Básicas, Salvador, Bahia, Brasil.

0000-0003-1195-0485

person Rafaela SCARIOT

Conceptualization (Equal) · Methodology (Equal) · Validation (Equal) · Visualization (Equal · Writing – review & editing (Equal)

schoolUniversidade Federal do Paraná, setor de Ciências da Saúde, Departamento de Estomatologia, Disciplina de Cirurgia Bucomaxilofacial, Curitiba, PR, Brasil.

0000-0002-4911-6413

person Luiz Evaristo Ricci VOLPATO

Conceptualization (Equal) · Data curation (Equal) · Formal analysis (Equal) · Investigation (Equal) · Methodology (Equal) · Project administration (Equal) · Resources (Equal) · Supervision (Equal) · Validation (Equal) · Visualization (Equal) · Writing – original draft (Equal) · Writing – review & editing (Equal)

schoolUniversidade de Cuiabá, Programa de Pós-Graduação em Ciências Odontológicas Integradas, Faculdade de Odontologia de Cuiabá, Cuiabá, MT, Brasil.

0000-0002-2969-1963

Corresponding address: Prof. Luiz Evaristo Ricci Volpato Av. Manoel José de Arruda, 3100 - Jardim Europa - 78065-700 - Cuiabá - MT - Brasil. Phone: 55-65-98114-5244 e-mail:
odontologiavolpato@uol.com.br

Editor: Ana Carolina Magalhães

Associate Editor: Renato Menezes Silva

Conflict of interest

The authors declare no conflict of interest.

Universidade de Cuiabá, Programa de Pós-Graduação em Ciências Odontológicas Integradas, Faculdade de Odontologia de Cuiabá, Cuiabá, MT, Brasil.

Universidade Estadual de Campinas, Departamento de Diagnóstico Oral e Programa de Pós-Graduação em Biologia Buco-Dental, Faculdade de Odontologia de Piracicaba, Piracicaba, SP, Brasil.

Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba, Programa de Pós-Graduação em Biologia Buco-Dental, Piracicaba, SP, Brasil.

Universidade Estadual de Montes Claros, Departamento de Odontologia, Clínica de Estomatologia, Montes Claros, MG, Brasil, e

Universidade Professor Edson Antônio Velano, Faculdade de Odontologia, Centro de Reabilitação de Anomalias Craniofaciais, Alfenas, MG, Brasil.

Escola Bahiana de Medicina e Saúde Pública, Departamento de Ciências Básicas, Salvador, Bahia, Brasil.

Universidade Federal do Paraná, setor de Ciências da Saúde, Departamento de Estomatologia, Disciplina de Cirurgia Bucomaxilofacial, Curitiba, PR, Brasil.

	Control	NSCL±P	NSCLO	NSCLP	NSCPO
	(n=881)	(n=801)	(n=233)	(n=568)	(n=273)
Sex
Male	421 (47.8%)	451 (56.3%)*	125 (53.6%)	326 (57.4%)*	105 (38.5%)
Female	460 (52.2%)	350 (43.7%)	108 (46.4%)	242 (42.6%)	168 (61.5%)
Ancestry
European	83.4%	82.7%	82.4%	82.7%	80.9%
African	14.8%	15.4%	15.9%	15.3%	16.6%
Amerindian	1.8%	1.9%	1.7%	2.0%	2.5%

Gene	SNP	Position	Allele	MAF	Call Rate	HWE (p-value)
WNT5A (chr 3)	rs566926	55486750	A/c	0.322	97.0%	0.72
FGFR1 (chr 8)	rs7829058	38474577	G/c	0.186	100.0%	0.07
BMP4 (chr 14)	rs11623717	53947414	A/g	0.359	98.5%	0.38
	rs17563	53950804	A/g	0.325	99.2%	0.10
	rs2071047	53951693	G/a	0.410	98.5%	0.78
	rs2761887	53958334	C/a	0.588	99.2%	0.51
GREM1 (chr 15)	rs16969681	32700910	C/t	0.197	99.9%	0.49
	rs16969816	32701444	G/a	0.324	99.8%	0.12
	rs16969862	32701800	A/c	0.222	99.6%	0.37
	rs1258763	32758222	C/t	0.459	99.5%	0.39
MMP2 (chr 16)	rs243836	55500324	G/a	0.464	98.8%	0.74
WNT3 (chr 17)	rs11653738	46809587	T/c	0.264	100.0%	0.49

	Control %	NSCL±P %	OR (95% CI)/ p-value	NSCLO %	OR (95% CI)/ p-value	NSCLP %	OR (95% CI)/ p-value	NSCPO %	OR (95% CI)/ p- value
Allele
A	77.1	73.9	Reference	71.7	Reference	74.9	Reference	76.2	Reference
C	22.9	26.1	1.18 (1.00-1.39)/0.03	28.3	1.32 (1.04-1.67)/0.01	25.1	1.12 (0.94-1.34)/0.18	23.8	1.05 (0.83-1.31)/0.68
Genotype
AA	59.6	55.4	Reference	52.5	Reference	56.7	Reference	59.6	Reference
AC	34.9	37.0	1.13 (0.92-1.40)/0.24	38.6	1.26 (0.92-1.73)/0.15	36.3	1.08 (0.86-1.37)/0.50	33.2	0.94 (0.70-1.27)/0.68
CC	5.5	7.6	1.47 (0.98-2.22)/0.05	9.0	1.87 (1.07-3.28)/0.03	7.0	1.32 (0.84-2.09)/0.21	7.2	1.36 (0.77-2.40)/0.29
Dominant (AA/AC+CC)	59.6/40.4	55.4/44.6	1.18 (0.97-1.44)/0.10	52.5/47.5	1.34 (1.00-1.81)/0.05	56.7/43.3	1.12 (0.89-1.39)/0.33	59.6/40.4	0.99 (0.75-1.32)/0.96
Recessive (AA+AC/CC)	94.5/5.5	92.4/7.6	1.40 (0.94-2.09)/0.09	91.0/9.0	1.70 (0.99-2.94)/0.06	93.0/7.0	1.28 (0.82-2.01)/0.27	92.8/7.2	1.39 (0.80-2.43)/0.25

	Control %	NSCL±P %	OR (95% CI)/ p-value	NSCLO %	OR (95% CI)/ p-value	NSCLP %	OR (95% CI)/ p-value	NSCPO %	OR (95% CI)/ p-value
Allele
A	76.8	72.6	Reference	71.5	Reference	73.0	Reference	73.5	Reference
C	23.2	27.4	1.25 (1.04-1.51)/0.01	28.5	1.32 (1.00-1.74)/0.04	27.0	1.22 (0.99-1.50)/0.05	26.5	1.19 (0.91-1.55)/0.18
Genotype
AA	59.6	5.1	Reference	51.5	Reference	53.7	Reference	56.1	Reference
AC	34.4	39.0	1.25 (0.98-1.60)/0.07	39.9	1.34 (0.93-1.93)/0.13	38.7	1.22 (0.93-1.60)/0.14	34.9	1.06 (0.74-1.50)/0.76
CC	5.9	7.9	1.45 (0.91-2.30)/0.11	8.6	1.64 (0.85-3.18)/0.15	7.6	1.36 (0.82–2.26)/0.24	9.0	1.56 (0.84–2.89)/0.19
Dominant (AA/AC+CC)	59.6/40.4	53.1/46.9	1.28 (1.02-1.62)/0.03	51.5/48.5	1.38 (0.98-1.95)/0.06	53.7/46.3	1.24 (0.96-1.60)/0.09	56.1/43.9	1.13 (0.81-1.57)/0.47
Recessive (AA+AC/CC)	94.1/5.9	92.1/7.9	1.32 (0.84-2.08)/0.22	91.4/8.6	1.46 (0.77-2.77)/0.25	92.4/7.6	1.25 (0.76-2.07)/0.37	91.0/9.0	1.52 (0.84-2.78)/0.18

	Control %	NSCL±P %	OR (95% CI)/ p-value	NSCLO %	OR (95% CI)/ p-value	NSCLP %	OR (95% CI)/ p-value	NSCPO %	OR (95% CI)/ p-value
Allele
A	77.7	77.8	Reference	72.5	Reference	80.0	Reference	82.9	Reference
C	22.3	22.2	0.99 (0.71-1.38)/0.98	27.5	1.32 (0.83-2.09)/0.23	20.0	0.86 (0.59-1.25)/0.45	17.1	0.72 (0.44-1.16)/0.17
Genotype
AA	59.5	62.1	Reference	55.0	Reference	65.2	Reference	68.4	Reference
AC	36.4	31.3	0.84 (0.56-1.28)/0.41	35.0	1.09 (0.59-2.02)/0.82	29.7	0.76 (0.48-1.21)/0.23	28.9	0.68 (0.38-1.22)/0.18
CC	4.1	6.6	1.49 (0.61-3.63)/0.35	10.0	2.57 (0.85-7.81)/0.09	5.1	1.11 (0.40-3.10)/0.82	2.6	0.61 (0.13-2.95)/0.51
Dominant (AA/AC+CC)	59.5/40.5	62.1/37.9	0.91 (0.61-1.36)/0.65	55.0/45.0	1.25 (0.70-2.23)/0.45	65.2/34.8	0.80 (0.51-1.24)/0.31	68.4/31.6	0.67 (0.38-1.18)/0.16
Recessive (AA+AC/CC)	95.9/4.1	93.4/6.6	1.58 (0.66-3.80)/0.30	90.0/10.0	2.49 (0.84-7.37)/0.11	94.9/5.1	1.22 (0.44-3.36)/0.70	97.4/2.6	0.69 (0.14-3.31)/0.63

	SNP1	SNP2	NH^a	betaH^b	NL^c	betaL^d	p-value^e	Perm. P-value^f
NSCL±P	rs566926 (WNT5A)	rs2071047 (BMP4)	1	13.357	1	-0.2479	2.13e-03	0.03
	rs566926 (WNT5A)	rs16969681 (GREM1)	1	0.6185	0	-	7.82e-03	0.05
	rs566926 (WNT5A)	rs16969862 (GREM1)	1	0.6231	0	-	8.14e-03	0.04
	rs566926 (WNT5A)	rs11623717 (BMP4)	1	0.8021	0	-	2.58e-02	0.21
	rs566926 (WNT5A)	rs1258763 (GREM1)	1	0.6355	1	-0.3667	4.57e-02	0.32
	rs566926 (WNT5A)	rs16969816 (GREM1)	1	0.5046	0	-	5.38e-02	0.33
	rs566926 (WNT5A)	rs243836 (MMP2)	0	-	1	-0.2047	6.77e-02	0.34
NSCLP	rs566926 (WNT5A)	rs7829058 (FGFR1)	2	0.3338	2	-0.5739	0.006	0.04
	rs566926 (WNT5A)	rs2071047 (BMP4)	1	11.687	1	-0.2821	0.01	0.12
	rs566926 (WNT5A)	rs16969681 (GREM1)	1	0.5504	0	-	0.03	0.17
NSCLO	rs566926 (WNT5A)	rs2071047 (BMP4)	1	16.480	0	-	0.001	0.02
	rs566926 (WNT5A)	rs2761887 (BMP4)	2	0.9896	1	-0.4462	0.002	0.05
	rs566926 (WNT5A)	rs243836 (MMP2)	0	-	1	-0.4675	0.009	0.09
	rs566926 (WNT5A)	rs16969681 (GREM1)	1	0.7645	0	-	0.01	0.10
	rs566926 (WNT5A)	rs16969862 (GREM1)	1	0.7203	1	- 0.5429	0.02	0.16
NSCPO	rs566926 (WNT5A)	rs2761887 (BMP4)	1	0.3755	2	-0.4430	0.003	0.04
	rs566926 (WNT5A)	rs2071047 (BMP4)	2	0.4912	0	-	0.01	0.17

[TFN1] * Statistically different from the control group at p<0.0005

[TFN2] NSCL±P: non-syndromic cleft lip with or without cleft palate; NSCLO: non-syndromic cleft lip only; NSCLP: non-syndromic cleft lip and palate; NSCPO: non-syndromic cleft palate only

Exploring the role of the WNT5A rs566926 polymorphism and its interactions in non-syndromic orofacial cleft: a multicenter study in Brazil