These double mutation cases included: case NTD_27 with CELSR2 p.Thr2026Met and DVL3 p.Asp403Asn, case NTD_39 with CELSR2 p.Arg2480Cys and FZD7 p.Leu383Gln, case NTD_56 with CELSR2 p.Arg2626Cys and FZD5 p.Trp242Leu, and case NTD_19 with CELSR1 p.Thr1086Met and VANGL1 p.Arg207His presented with spina bifida. This evidence concerns the gene FZD5 and spina bifida.