Table 2.
Sample | Used for | Sample type/ Predicted pathogenicity | TET3 amino acid changes | CADD score | GnomAD alleles | Inheritance | In catalytic domain? | Evidence supporting pathogenicity | Evidence against pathogenicity | Epi-signature pathogenicity prediction |
---|---|---|---|---|---|---|---|---|---|---|
1a | Signature discovery | TET3 (BA); path | p.Phe1072Cysc; p.Ala1076Thrc | 28.2; 25.9 | 0; 0 | AR, Cpd het | Yes; Yes | Both low TET activityi; CADD; gnomAD; inheritance; in cat domain | None | Both path |
2a | Signature discovery | TET3 (BA); path | p.Val908Leu; p.Val908Leuc | 27 | 0 | AR, Hom | Yes | Low TET activityi; CADD; gnomAD; inheritance; in cat domain | None | Path |
3a | Signature discovery | TET3 (BA); path | p.Val908Leu; p.Val908Leuc | 27 | 0 | AR, Hom | Yes; Yes | Low TET activityi; CADD; gnomAD; inheritance; in cat domain | None | Path |
4a | Signature discovery | TET3 (MA); path | p.His1660Profs*52 | NA | 0 | AD, inherited | Yes | Predicted LOF; gnomAD; inheritance; in cat domain | None | Path |
5a | Signature discovery | TET3 (MA); path | p.His1660Profs*52 | NA | 0 | Unkn | Yes | Predicted LOF; gnomAD; in cat domain | None | Path |
6a | Signature discovery | TET3 (MAb); path | p.Val1089Metc; p.Arg752Cysc | 29.1; 23.6 | 0; 29 (0 hom) | Initially AR; now ADb | Yes; No | p.Val1089Met low TET activityi; CADD; gnomAD; in cat domain. p.Arg752Cys CADD | p.Val1089Met none; p.Arg752Cys nl TET activityi; gnomAD; not in cat domain | Path/Intermed |
7 | Signature validation | Family ctld; NA | Family variant absent (Sanger seq) | NA | NA | NA | NA | NA | NA | Non-path |
8 | Signature validation | Family ctle; NA | Family variant absent (Sanger seq) | NA | NA | NA | NA | NA | NA | Non-path |
9 | Signature validation | Family ctlf; NA | Family variant absent (Exome seq) | NA | NA | NA | NA | NA | NA | Non-path |
10 | Signature validation | Family ctlg; NA | Family variants absent (Sanger, exome seq) | NA | NA | NA | NA | NA | NA | Non-path |
11 | Signature validation | TET3 (MA); benign | p.Arg752Cysc | 23.6 | 29 (0 hom) | Unkn | No | CADD | Nl TET activityi; gnom-AD; not in cat domain | Intermed |
12a | Signature validation | TET3 (MA); path | p.Gln1695* | 44 | 0 | AD, de novo | Yes | Predicted LOF; CADD; gnomAD; inheritance; in cat domain | None | Path |
13a | Signature validation | TET3 (MA); path | p.Arg1034* | 38 | 0 | AD, de novo | Yes | Predicted LOF; CADD; gnomAD; inheritance; in cat domain | None | Path |
14a | Signature validation | TET3 (MA); path | p.Val1089Metc | 29.1 | 0 | Unkn | Yes | Low TET activityi; CADD; gnomAD; in cat domain. | None | Path |
15a | Signature validation | TET3 (MA); path | p.Val908Leuc | 27 | 0 | Unkn | Yes | Low TET activityi; CADD; gnomAD; in cat domain | None | Path |
16a | Signature validation | TET3 (MA); path | p.Ala1076Thrc | 25.9 | 0 | Unkn | Yes | Low TET activityi; CADD; gnomAD; in cat domain | None | Path |
17 | Testing | TET3 (BA) VUS; unkn | p.Pro495Ser; p.Val1295Ile | 7.9; 19 | 13 (0 hom); 127 (0hom) | AR, Cpd het | No; Yes | p.Pro495Ser none p.Val1295Ile in cat domain | Both CADD; gnomAD; p.Pro495Ser not in cat domain | Both likely benign |
18 | Testing | TET3 (BA) VUS; unkn | p.Gly1505Arg; p.Trp1746Ser | 24.5; 29.3 | 3 (0 hom); 61 (0 hom) | AR, Cpd het | Yes; Yes | CADDs; both in cat domain | GnomAD; both present in unaffected sib | Both likely benign |
19 | Testing | TET3 (MA) VUS; unkn | p.Pro495Ser | 7.9 | 13 (0 hom) | Unkn | No | None | CADD; gnomAD; not in cat domain | Likely benign |
20 | Testing | TET3 (MA) VUS; unkn | p.Val1295Ile | 19 | 127 (0 hom) | Unkn | Yes | In cat domain | CADD; gnomAD | Likely benign |
21a | Testing | TET3 (MA) VUS; unkn | p.Arg911Gln | 27.1 | 0 | AD, de novo | Yes | CADD; gnomAD; inheritance; in cat domain | None | Likely path |
22a | Testing | TET3 (MA) VUS; unkn | p.Arg1683His | 31 | 0 | AD, de novo | Yes | CADD; gnomAD; inheritance; in cat domain | None | Likely path |
23 | Testing | TET3 (MA) VUS; unkn | p.Gly1505Arg | 24.5 | 3 (0 hom) | Unkn | Yes | CADD; in cat domain | GnomAD; this variant, p.Trp1746Ser in unaffected sib | Likely benign |
24 | Testing | TET3 (MA) VUS; unkn | p.Trp1746Ser | 29.3 | 61 (0 hom) | Unkn | Yes | CADD; in cat domain | GnomAD; this variant, p.Gly1505Arg in unaffected sib | Likely benign |
25a | Testing | TET3 (MA) VUS; unkn | p.Thr680Tyrfs*26h | NA | 0 | Unkn | No | Predicted LOF; gnomAD; inheritance-segregates with phenotype in family | None | Likely path |
26a | Testing | TET3 (MA) VUS; unkn | p.Thr680Tyrfs*26h | NA | 0 | AD, inherited | No | Predicted LOF; gnomAD; inheritance-segregates with phenotype in family | None | Likely path |
27a | Testing | Episign screen; unkn | p.Cys246* | 34 | 0 | AD, inherited | No | Predicted LOF; CADD; gnomAD; inherited from mosaic mother | None | Likely path |
BA bi-allelic, MA mono-allelic, path pathogenic, non-path non-pathogenic, NA not applicable, unkn unknown, AR autosomal recessive, Cpd het compound heterozygote, Hom homozygous, AD autosomal dominant, LOF loss-of-function, cat catalytic, nl normal, seq sequencing, sib sibling, CADD score combined annotation-dependent depletion score (https://cadd.gs.washington.edu/); gnomAD browser is at https://gnomad.broadinstitute.org/.
aTET3-deficient pathogenic samples used to identify the final DNA methylation episignature.
bConsidered mono-allelic because only the c.3265G>A (p.Val1089Met) variant reduced catalytic activity in vitro, suggesting pathogenicity; c.2254C>T (p.Arg752Cys) did not reduce catalytic activity in vitro, suggesting non-pathogenicity (ref. 8).
cVariants previously tested in our in vitro assay (ref. 8).
dUnaffected daughter of Sample 5 and sister of Sample 4.
eUnaffected son of 25 and brother of 26.
fUnaffected mother of Sample 4.
gUnaffected sister of Sample 6 and daughter of Samples 11 and 14.
hPreviously considered VUS because mother was not thought to be affected; upon further evaluation, she was noted to be affected (see text for details).
iTET activity based on in vitro assay performed and reported in reference8.