********** MammakarzinomMammakarzinom DR. SENCKENBERGISCHESDR. SENCKENBERGISCHES PatientPatient name name DiagnosisDiagnosis R PATHOLOGIER PATHOLOGIE PatientPatient 1D 1D Testl234Testl234 ICD-10-CMICD-10-CM code code ;;:;;;:; spsp CaseCase 1D 1D EU001930EU001930 MeSHMeSH ID/term ID/term D001943D001943 (Breast (Breast Neoplasms) Neoplasms) W1ldlabW1ldlab DateDate of birth of birth AdditionalAdditional MeSH MeSH IDs IDs - - SexSex FemaleFemale SampleSample type type FFPEFFPE LabtestLabtest VCF-completeVCF-complete import import (paired) (paired) GeneralGeneral dataset dataset 1D 1D105685598965 105685598965 EthnicityEthnicity EUR EUR TumorTumor cellularity cellularity 100% 100% OrganizationalOrganizational unit unit SIPV2TESTSIPV2TEST CVICVI dataset dataset 1D 1D 105685598965105685598965 CountryCountry DE DE SoftwareSoftwareversionversion 4.3.34.3.3 MutationalMutational status status of commonly of commonly mutated mutated genes genes in the in the patient patient disease disease ATMATM BRCAlBRCAl BRCA2BRCA2 CDK4CDK4 CDKN2ACDKN2A CHEK2CHEK2 EGFREGFR ERBB2ERBB2 ESRlESRl PALB2PALB2 PIK3CAPIK3CA notnot notnot notnot notnot notnot notnot notnot notnot notnot notnot 1 SNV1 SNV identifiedidentified identifiedidentified identifiedidentified identifiedidentified identifiedidentified identifiedidentified identifiedidentified identifiedidentified identifiedidentified identifiedidentified PTENPTEN TP53TP53 notnot notnot identifiedidentified identifiedidentified SUMMARYSUMMARY OverviewOverview of potential of potential treatment treatment impacts impacts OverviewOverview of prognostic of prognostic and and diagnostic diagnostic findings findings ClinicalClinical trials trials found found 3 3Effective Effective 1 1lneffective lneffective o Safetyo Safety O OPrognostic Prognostic O ODiagnostic Diagnostic 6 6Trials Trials VAF/CopyVAF/Copy PotentialPotential impact impact TreatmentTreatment DrugDrug approval approval BiomarkerBiomarker BiomarkerBiomarker score score TrialsTrials NumberNumber AlpelisibAlpelisib ApprovedApproved PIK3CAPIK3CA ClinicallyClinically EffectiveEffective 66.67%66.67% 4 4 FulvestrantFulvestrant ApprovedApproved p.E545Ap.E545A (SNV) (SNV) r;,r;, Approved Approved -- �� PIK3CAPIK3CA EffectiveEffective EverolimusEverolimus Approved*Approved* 66.67%66.67% 2)2) Preclinical Preclinical 2 2 p.E545Ap.E545A (SNV) (SNV) -- PIK3CAPIK3CA EffectiveEffective SirolimusSirolimus OtherOther 66.67%66.67% 2)2) Preclinical Preclinical 0 0 p.E545Ap.E545A (SNV) (SNV) -- PIK3CAPIK3CA lneffectivelneffective TrastuzumabTrastuzumab Approved*Approved* 66.67%66.67% 4)4)ClinicalClinical p.E545Ap.E545A (SNV) (SNV) 1 1 �� * the* the drug drug is approved is approved for forthe the cancer cancer type type but but either either none none of theof the currently currently approved approved biomarkers biomarkers for forthis- this drug- drug were were identified, identified, or anor anapproved approved resistance resistance biomarkerbiomarker for forthe the drug drug was was identified identified in this in this patient. patient. Therefore, Therefore, the the drug drug labe[ labe[ may may not not cover cover the the analyzed analyzed patient; patient; VAF VAF = Variant = Variant allele allele frequency frequency BiomarkerBiomarker score: score: AMP AMP score score and and CVI CVI score. score. Clinically Clinically approved: approved: Approved Approved biomarker biomarker (by (bythe the FDA, FDA, EMA, EMA, or NCCN)or NCCN) to predictto predict a specific a specific effect effectin thein the patient's patient's disease.disease. Clinical: Clinical: Not Not yet yetapproved approved biomarker biomarker for forthe the patient's patient's disease. disease. Observed Observed in clinical in clinical studies studies as aas potential a potential biomarker biomarker to predictto predict a specific a specific effect effectof theof the drug.drug. Preclinical: Preclinical: This This biomarker biomarker has has not not yet yetbeen been observed/tested observed/tested in patients in patients to predict to predict a specific a specific effect effectof theof the drug. drug. lt is lt supported is supported by preclinicalby preclinical evidence evidence or or translationaltranslational data. data. YouYou can can find find more more details details on theon the biomarker biomarkerscorescore (AMP (AMP and and CVI CVI score) score) in the in the glossary. glossary. TheodorTheodor Stern Stern Kai 7, Kai 7, SignedSigned by: by: MolecularMolecular Health Health GmbH GmbH 6059060590 Frankfurt Frankfurt ReportReportdate: date: 08 Mar 08 2021 Mar 202116:5816:58 (UTC+Ol:00) (UTC+Ol:00) Kurfürsten-AnlageKurfürsten-Anlage 21, 21, ReportReportversion: version: 3 3 6911569115 Heidelberg Heidelberg OrderOrder date: date: 08 Mar 08 2021 Mar 2021 Page Page1 of 13 1 of 13 Patient name ***** Diagnosis Mammakarzinom DR. SENCKENBERGISCHESDR. SENCKENBERGISCHES Patient name***** Diagnosis Mammakarzinom R PATHOLOGIER PATHOLOGIE PatientPatient 1D 1D Testl234Testl234 ICD-10-CMICD-10-CM code code ;;:;;;:; spsp CaseCase 1D 1D EU001930EU001930 MeSHMeSH ID/term ID/term D001943D001943 (Breast (Breast Neoplasms) Neoplasms) W1ldlabW1ldlab DateDate of birth of birth AdditionalAdditional MeSH MeSH IDs IDs - - Sex Female Sample type FFPE Labtest VCF-complete import (paired) General dataset 1D 105685598965 BIOMARKEREthnicity EUR DETAILS Tumor cellularity 100% Organizational unit SIPV2TEST CVI dataset 1D 105685598965 Country DE Software version 4.3.3 Mutational status of commonly mutated genes in the patient disease PIK3CA p.E545A (SNV) 1 ATM BRCAl BRCA2 CDK4 CDKN2A CHEK2 EGFR ERBB2 ESRl PALB2 PIK3CA not not not not not not not not not not 1 SNV ••identified identified identified identified identified identified identified identified identified identified PIK3CA is the catalytic subunit of the lipid phosphoinositide-3-kinase (Pl3K) that activates the Pl3K/AKT signaling pathway to promote cell proliferation and survival.PTEN This variantTP53 strongly activates the downstream pathway in preclinical settings. Alpelisib plus fulvestrant is indicated for this variant for the treatment of hormonenot receptor-positive,not H ER2-negative advanced breast cancer. The HER2-positive tumor of a patient with this variant was resistance to trastuzumab. Althoughidentified not specifically identified tested for this mutation, preclinical models with activating variants are sensitive to the mTOR inhibitors everolimus and sirolimus and have reduced sensitivity to trastuzumab. A [arger study showed that HER2-positive patients with an activating PIK3CA mutation had improved progression-free survival with the addition of everolimus to trastuzumab treatment. PubMedSUMMARY 1D 21358673,28382169,27091708,31091374,23092874 Overview of potential treatment impacts Overview of prognostic and diagnostic findings Clinical trials found 3 Effective 1 lneffective o Safety O Prognostic O Diagnostic 6 Trials Potential impact Treatment Drug approval Biomarker score Clinically Effective Alpelisib + Fulvestrant Approved, ApprovedVAF/Copy Potential impact Treatment Drug approval Biomarker Biomarker score r;, ApprovedTrials Number - Effective EverolimusAlpelisib Approved Approved*PIK3CA Clinically2, Preclinical Effective 66.67% 4 Fulvestrant Approved p.E545A (SNV) r;, Approved -� Effective - 2' Preclinical Sirolimus OtherPIK3CA Effective Everolimus Approved* 66.67% 2) Preclinical 2 p.E545A (SNV) - lneffective Trastuzumab Approved* - Clinical PIK3CA Effective Sirolimus Other 66.67% • 2) Preclinical4, 0 p.E545A (SNV) � * the drug is approved for the cancer type but either none of the currently approved biomarkers for this drug were- identified, or an approved resistance biomarker for the drug was identified in this patient. Therefore, the drug labe[ may not cover the analyzed patient.- PIK3CA ) Clinical Biomarkerlneffective score: AMP score andTrastuzumab CVI score. Clinically Approved* approved: Approved biomarker (by the66.67% FDA, EMA, or NCCN) to predict1 a specific4 effectin the patient's p.E545A (SNV) � disease. Clinical: Not yet approved biomarker for the patient's disease. Observed in clinical studies as a potential biomarker to predict a specific effectof the drug.* the Preclinical: drug is approved This biomarker for the has cancer not yettype been but observed/testedeither none of the in patients currently to approved predict a specificbiomarkers effect forof thisthe- drug.drug ltwere is supported identified, by or preclinical an approved evidence resistance or translationalbiomarker data. for the drug was identified in this patient. Therefore, the drug labe[ may not cover the analyzed patient; VAF = Variant allele frequency YouBiomarker can find more score: details AMP on score the biomarker and CVI score.score Clinically (AMP and approved: CVI score) in Approved the glossary. biomarker (by the FDA, EMA, or NCCN) to predict a specific effectin the patient's disease. Clinical: Not yet approved biomarker for the patient's disease. Observed in clinical studies as a potential biomarker to predict a specific effectof the drug. Preclinical: This biomarker has not yet been observed/tested in patients to predict a specific effectof the drug. lt is supported by preclinical evidence or translational data. You can find more details on the biomarker score (AMP and CVI score) in the glossary. TheodorTheodor Stern Stern Kai 7, Kai 7, SignedSigned by: by: MolecularMolecular Health Health GmbH GmbH 6059060590 Frankfurt Frankfurt ReportReportdate: date: 08 Mar 08 2021 Mar 202116:5816:58 (UTC+Ol:00) (UTC+Ol:00) Kurfürsten-AnlageKurfürsten-Anlage 21, 21,