Case1: Male, 40 years old, French

DiseaseAssertion: Glanzmann thrombasthenia

FamilyInfo: Mild bleeding symptoms have been reported in his grandmother and a first cousin, but they were not available for further study

ParentalGenotype: The mother is asymptomatic; however, genetic testing confirmed her to be heterozygous for the GT mutation (No information on father's genetic information for this mutation)

CasePresentingHPOs: HP:0000225

(Gingival bleeding)

CaseHPOFreeText: Very mind bleeding

CaseNotHPOs: HP:0000421, HP:0000978

(Epistaxis, bruising)

CaseNotHPOsFreeText: N/A

ProteinExpression: The surface expression of GPIIb-IIIa was about 20% of normal, whereas GPIb (93%) was normally expressed (See Figure 1A)

PlateletAggregation: Platelet aggregation was markedly impaired in response to physical agonists including: ADP, arachidonic acid, epinephrine, and collagen; however, ristocetin induced normal aggregation

CasePreviousTesting: All 30 exons and immediate flanking sequences of the GPIIb gene, and all 15 exons and immediate flanking sequences of the GPIIIa gene, were amplified by PCR.

Gene: ITGB3

Variant: NM_000212.2:c.1756T>C (p.Cys586Arg)

LegacyVariant: (p.Cys560Arg)

ClinVar: N/A

CAiD: CA400032477

gnomAD: N/A

Case1: (Patient RM) Male, age and ethnicity not specified

DiseaseAssertion: Glanzmann thrombasthenia

FamilyInfo: N/A

ParentalGenotype: Mother: heterozygous for (p.Arg724Ter)

CasePresentingHPOs:

CaseHPOFreeText: HP:0000978 (Easy bruising)

CaseNotHPOs: Recurrent mucosal bleeding

CaseNotHPOFreeText: N/A

ProteinExpression: Flow cytrometric analysis revealed reduced levels of alphaIIbbeta3 on the surface of RM platelets (See Fig 1B)

PlateletAggregation: Ristocetin induced agglutination was normal; however, platelets failed to aggregate in response to all physiological agonists including ADP, collagen, epinephrine, thrombin, and arachidonic acid

CasePreviousTesting: Direct DNA sequence analysis of PCR amplified ITGA2B and ITGB3 genes from patient RM's genomic DNA was performed. All exons, including the immediate intron-exon junctions of ITGA2B and ITGB3 were amplified using the PCR primer pairs originating in the intron sequence flanking each exon. The resulting PCR fragments were purified and subjected to direct cycle sequencing

Gene: ITGB3

Variant: NM_000212.2:c.2248C>T (p.Arg750Ter) and NM_000212.2:c.1791del (p.Asn597Lysfs*72)

LegacyVariant: (p.Arg724Ter)

ClinVar: 13564

CAiD: CA123246 and CA8623358

gnomAD: MAF in the African population is 0.0006008 https://gnomad.broadinstitute.org/variant/17-45384950-C-T

PreviouslyPublished: PMID: 15701721

Case1: (Patient RM), Male, 73 years old, from Southwest of France

DiseaseAssertion: Glanzmann thrombasthenia

FamilyInfo: Other family members were his mother (M.M), his son (J.M.M), and his sister (A.M)

ParentalGenotype: Mother is heterozygous for the mutation

CasePresentingHPOs: HP:0000421 (Epistaxis)

CaseHPOFreeText: Life-long history of mucocutaneous bleeding

CaseNotHPOs: N/A

ProteinExpression: Flow cytometric analysis shows a reduced aIIbb3 expression in the platelets of the proband and other family members (See Fig. 6)

Platelet Aggregation: Absent platelet aggregation in response to ADP, collagen, arachidonic acid and thrombin receptor activated peptide. Ristocetin induced platelet agglutination was normal

CasePreviousTesting: Screening of the aIIb and b3 genes by PCR-SSCP revealed aheterozygous mutation at position 685 in exon 5 of the b3 gene leading to a 1 96Leu to Pro substitution.

Gene: ITGB3

Variant: NM_000212.2:c.665T>C (p.Leu6656Pro)

LegacyVariant: (p.Leu196Pro)

ClinVar: N/A

CAiD: CA291224887

gnomAD MAF in the European (non-Finnish) population is 0.00006482 https://gnomad.broadinstitute.org/variant/17-45363676-T-C

Case2: Male, 9 years old, Japanese

DiseaseAssertion: Macrothrombocytopenia

FamilyInfo: Information on mother is given (minimal other family information)

ParentalGenotype: Mother: ITGA2B (Phe993del heterozygous)

CasePresentingHPOs: N/A

CaseHPOFreeText: N/A

CaseNotHPOs: N/A

CaseNotHPOsFreeText: N/A

ProteinExpression: Surface expression of αIIbβ3 in platelets of the patient and his mother was about 80% compared to control subjects with an increase of CD42b expression

PlateletAggregation: N/A

CasePreviousTesting: The entire coding sequence of exons and exon-intron boundaries of ITGA2B and ITGB3 and the whole coding regions of ITGA2B and ITGB3 cDNA obtained from platelets were sequenced as previously described. In the case of ITGA2B p.Phe993del, PCR products obtained from amplification of exon 30 of ITGA2B from the patient's DNA were subcloned to pCR2.1-TOPO and sequence analysis was performed

Gene: ITGA2B

Variant: NM_000419.3:c.3070_3072del (p.Phe1024del)

LegacyVariant: p.Phe993del legacy nomenclature

ClinVar: N/A

CAiD: CA8602460

gnomAD: N/A

Case3: Female, 8 years old, Japanese

DiseaseAssertion: Thrombocytopenia

FamilyInfo: Information on mother and grandfather available in Table 1(minimal information on other family members)

ParentalGenotype: Mother: ITGB3 (p.Asp621_Glu660del heterozygous) See Table 1

CasePresentingHPOs: HP:0000978 (Bruising)

CaseHPOFreeText: Subaponeurotic hematoma

CaseNotHPOs: N/A

CaseNotHPOsFreeText: N/A

ProteinExpression: Surface expression levels of αIIbβ3 in platelets of patient were decreased to around 65% of normal subjects with increase of CD42b (See Table 1)

PlateletAggregation: N/A

CasePreviousTesting: Sequence analysis of DNA revealed the patient's mutation. RT-PCR was also performed. The entire coding sequence of exons and exon-intron boundaries of ITGA2B and ITGB3 and the whole coding regions of ITGA2B and ITGB3 cDNA obtained from platelets were sequenced.

Gene: ITGB3

Variant: NM_000212.2:c.2134+1G>A (p.Asp621_Glu660del)

LegacyVariant: N/A

ClinVar: N/A

CAiD: CA400033344

gnomAD: N/A

Case1: Female, 21 years old, Japanese (Named Osaka-12 in this study)

DiseaseAssertion: Glanzmann thrombasthenia

FamilyInfo: N/A

ParentalGenotype: The substitution was inherited from her father. The molecular genetic defect inherited from her mother remains determined.

CasePresentingHPOs: N/A

CaseHPOFreeText: Moderate mucocutaneous bleeding in childhood

CaseNotHPOs: N/A

CaseNotHPOsFreeText: N/A

ProteinExpression: Flow cytometry showed revealed a significant reduction in the expression of αIIbβ3 on Osaka-12 platelets

PlateletAggregation: Absent platelet aggregation in response to ADP, epinephrine, and collagen but a normal response to ristocetin

CasePreviousTesting: To identify the molecular genetic defect for the proband of interest, all coding regions of ITGA2B and ITGB3 cDNA amplified by RT-PCR were analyzed

Gene: ITGA2B

Variant: NM_000419.3:c.520T>C (p.Tyr174His)

LegacyVariant: (p.Tyr143His)

ClinVar: N/A

CAiD: CA399805805

gnomAD: N/A

Case1: Male, French, age not specified

DiseaseAssertion: Glanzmann thrombasthenia

FamilyInfo: Son was heterozygous carriers of the p.Leu196Pro mutation (Sister died from nasal bleeding when she was a child)

ParentalGenotype: Mother is heterozygous for p.Leu196Pro mutation (Father not alive)

CasePresentingHPOs: HP:0000421 (Epistaxis)

CaseHPOFreeText: Lifelong history of mucocutaneous bleeding

CaseNotHPOs: N/A

CaseNotHPOsFreeText: N/A

ProteinExpression: Expression showed 8% of normal platelet content for αIIbβ3

PlateletAggregation: Absent aggregation to physiologic agonists

CasePreviousTesting: Screened exons and splice sites of ITGB3 and ITGA2B genes of patient by single-strand conformation polymorphism analysis of PCR products.

Gene: ITGB3

Variant: NM_000212.2:c.665T>C (Leu222Pro) and NM_000212.2:c.1871G>A (p.Cys624Tyr)

LegacyVariant: p.Leu196Pro and p.Cys598Tyr

ClinVar: N/A

CAiD: CA291224887 and CA400032726

gnomAD: N/A

Case3: Gender or age not specified, Thai ethnicity

DiseaseAssertion: Glanzmann thrombasthenia

FamilyInfo: Parental information available in Table 2

ParentalGenotype: Father: p.Arg782Ter/WT and Mother: Unavailable

Note: No parental consanguinity

CasePresentingHPOs: N/A

CaseHPOFreeText: The article does not mention specific bleeding phenotypes for each of the 4 probands in the article, rather, there is a general statement that says: "The common presentations included recurrent epistaxis, bleeding in oral cavity, and easy-bruising spontaneously or after trauma"

CaseNotHPOs: N/A

CaseNotHPOsFreeText: N/A

ProteinEpression: General statement from the article that states that flow cytometry to assess the surface GPIIb/IIIa complex showed reduced expression for all probands.

PlateletAggregation: The platelet aggregation test was typically defective with all physiological agonists except ristocetin, for all patients.

CasePreviousTesting: Whole exome sequencing was performed on the 30 exons of ITGA2B and 15 exons in ITGB3. PCR and direct sequencing were then performed in the DNA samples of the patients and their available parents to verify the presence of the identified mutations.

Gene: ITGA2B

Variant: NM_000419.3:c.2344C>T (p.Arg782Ter) and NM_000419.3:c.2915dup (p.Leu973Alafs*63)

LegacyVariant: N/A

ClinVar: N/A

CAiD: CA399796586 (p.Arg782Ter) and CA8602531 (p.Leu973Alafs*63)

gnomAD: MAF for (p.Leu973Alafs*63) in East Asian population 0.0001088 http://gnomad.broadinstitute.org/variant/17-42452054-C-CG