or Liquid Biopsy

Accepted Manuscript Lung Cancer Patients with Germline Mutations detected by next generation sequencing and/or liquid biopsy Takehito Shukuya, MD, PhD, Sandipkumar Patel, MD, Kate Shane-Carson, MS, LGC, Kai He, MD, PhD, Erin M. Bertino, MD, Konstantin Shilo, MD, Gregory A. Otterson, MD, David P. Carbone, MD, PhD PII:

S1556-0864(17)32764-8

DOI:

10.1016/j.jtho.2017.09.1962

Reference:

JTHO 747

To appear in:

Journal of Thoracic Oncology

Received Date: 18 August 2017 Revised Date:

25 September 2017

Accepted Date: 26 September 2017

Please cite this article as: Shukuya T, Patel S, Shane-Carson K, He K, Bertino EM, Shilo K, Otterson GA, Carbone DP, Lung Cancer Patients with Germline Mutations detected by next generation sequencing and/or liquid biopsy, Journal of Thoracic Oncology (2017), doi: 10.1016/j.jtho.2017.09.1962. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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Lung Cancer Patients with Germline Mutations detected by next

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generation sequencing and/or liquid biopsy

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Takehito Shukuya MD, PhD1, Sandipkumar Patel MD1, Kate Shane-Carson, MS, LGC2, Kai

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He MD, PhD1, Erin M. Bertino MD1, Konstantin Shilo MD3, Gregory A. Otterson MD1,

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David P. Carbone MD, PhD1

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Columbus, Ohio, USA

Division of Medical Oncology, Department of Internal Medicine, The Ohio State University,

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Columbus, Ohio, USA

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Columbus, OH, USA

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Division of Human Genetics, Department of Internal Medicine, The Ohio State University,

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Thoracic Pathology Division, Department of Pathology, The Ohio State University,

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Correspondence to: David P. Carbone, MD, PhD, Division of Medical Oncology, Department

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of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA. Phone:

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+1-614-685-4479, FAX: +1-614-366-1969, E-mail: [email protected]

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Key words: lung cancer; germline mutation; next generation sequencing; liquid biopsy 1

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Funding sources

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This work was supported by The Lilly Oncology Fellowship from The Japanese Respiratory

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Society, alumni scholarship from the Juntendo University School of Medicine, a research

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fellowship from Uehara Memorial Foundation, Pelotonia postdoctoral fellowship (to TS) and

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the Blue Beautiful Skies Fund (to DPC).

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Conflicts of interest

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TS reports grants and personal fees from Chugai Pharmaceutical, grants from Boehringer

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Ingelheim, personal fees from AstraZeneca, Ono Pharmaceutical, Nichi-iko Pharmaceutical,

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Sanofi, and Pfizer, outside the submitted work. KSC reports grants from Addario Lung

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Cancer Foundation, outside the submitted work. GAO reports grants and other from Pfizer,

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and Genentech, grants from Pfizer, BMS, Ignyta, and Merck, other from Novartis, and

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Amgen, outside the submitted work. DPC reports personal fees from Abbvie, Adaptimmune,

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Amgen, Ariad, AstraZeneca, Bayer Health Care, Biocept, Biothera, Boehringer Ingelheim,

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Cancer

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Genentech/Roche, Genoptix, GlaxoSmithKline, Gritstone, Guardant Health, ImmuneDesign,

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Janssen Diagnostics, MedImmune, Merck, MSD, Novartis, Palobiofarma, Peregrine

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Pharmaceuticals, Inc., Pfizer, prIME Oncology, Stemcentrx, Synta Pharmaceuticals Corp.,

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Support

Community,

Celgene,

Clovis

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Oncology,

Foundation

Medicine,

ACCEPTED MANUSCRIPT Takeda, Teva Pharmaceuticals, Verastem, grants and personal fees from Bristol

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Myers-Squibb, outside the submitted work.

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ACCEPTED MANUSCRIPT Case 1 is a 50-year-old female never smoker with recurrent small-cell lung cancer

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who visited our institution to discuss further treatment plan in 2015. She had a family history

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of pancreatic cancer in a sibling and leukemia in another sibling. She underwent left upper

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lobe wedge resection of a primary lesion, adjuvant cisplatin plus etoposide, and topotecan at

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another institution. She had a past history of breast cancer. which was diagnosed in 2006.

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She underwent lumpectomy, and pathological findings from her breast cancer showed it to be

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ER positive, PR negative, and HER2 negative. Postoperatively, she was treated with adjuvant

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chemotherapy and radiotherapy followed by Tamoxifen. In 2010, she had an isolated lung

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lesion, and underwent left upper lobe lobectomy after needle biopsy. Final pathology showed

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low grade, moderately differentiated adenocarcinoma, ER + (15%), PR -, HER2 IHC 1+,

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HER2 FISH -, CK AE1/AE3 +, CK7 +, mammaglobin +, CK20 -, TTF1 -, and was consistent

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with metastatic breast carcinoma. She again received adjuvant chemotherapy. In 2015, CT

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scan showed 18x10x16 mm subpleural nodule in the left lower lung, and she underwent

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wedge resection. Final pathology on this specimen showed small cell lung cancer,

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synaptophysin +, CD56 +, TTF-1 +, HMWK focally +, CK5 -, GATA - (Figure 1). Both the

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small cell lung cancer specimen and metastatic breast cancer specimen were sent for NGS

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testing (FoundationOneTM), and TP53 Y236* mutation and PARK2 Q347* mutation were

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detected in both specimens (Table 1) [1]. Germline mutations in TP53 and PARK2 were

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suspected since these same mutations were present in both tumor specimens, and allele

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ACCEPTED MANUSCRIPT frequencies for these were requested. Allele frequencies of the TP53 Y236* mutation and the

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PARK2 Q347* mutation in the small cell lung cancer specimen were 83% and 47%,

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respectively. She was recommended to see genetics and to undergo germline genetic testing,

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but she declined. As exactly the same mutations in TP53 and PARK2 were detected in two

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separate cancer specimens (both the metastatic breast cancer specimen and the primary lung

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cancer specimen), and their allele frequencies were significantly increased in the lung

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specimen, we clinically diagnosed the patient with germline mutations in TP53 and PARK2.

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Targeted therapy directed against FGFR2 amplification was planned in a clinical trial setting,

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but unfortunately she did not meet the eligibility criteria of the clinical trial due to elevated

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liver enzymes.

Case 2 is a 34-year-old female former smoker with advanced lung adenocarcinoma

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(metastatic pleural effusion) who was referred to our institution for further treatment in 2016.

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She received pleural catheter placement, four cycles of carboplatin plus pemetrexed followed

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by two cycles of maintenance pemetrexed, and four doses of nivolumab. A blood sample was

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sent for cfDNA testing (FoundationACTTM) and MET 3028+2T>C (splice site mutation) and

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BRCA2 L3061* mutations were detected (Table 2) [1]. Allele frequencies for these mutations

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were 0.19% and 50.7%, respectively, and a germline mutation in BRCA2 was suspected. She

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was referred to genetics, and underwent testing for mutations in the BRCA2 gene. The result

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showed that she had a germline BRCA2 L3061* mutation. Of note, this patient had no

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ACCEPTED MANUSCRIPT personal or family history of breast or ovarian cancer that would have otherwise prompted

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germline genetic testing of the BRCA2 gene. She received crizotinib, and had a partial

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response.

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These two cases suggest that practicing oncologists should be aware that detecting

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significant germline mutations in lung cancer patients is possible with currently available and

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routine technologies.

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Germline mutations predisposing to lung cancer are rare, but germline mutations in

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EGFR, HER2, BRCA2, CDKN2A, BAP1, SFTPA2, and PARK2 are associated with an

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increased risk of lung cancer [2]. We retrospectively investigated 3,869 lung cancer patients

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seen at our institution between February 2012 and January 2017, and seven patients including

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two patients presented here were found to have potentially predisposing germline mutations

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(Three had BRCA2 germline mutations, two had germline TP53 mutations [of which one

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patient also had a PARK2 mutation], one had a BRCA1 mutation, and one had an EGFR

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T790M mutation.).

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Introduction of NGS technology and liquid biopsies to clinical practice can raise the

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probability of detecting germline mutations in lung cancer patients [1, 3]. As in our patient

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from Case 2, this can have implications for screening and preventative measures for cancers

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besides lung cancer, as well as implications for cancer risks for other relatives. Clinicians

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should be alert to the potential existence and importance of germline mutations in their lung 6

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cancer patients.

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Acknowledgments

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This work was supported by The Lilly Oncology Fellowship from The Japanese Respiratory

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Society, alumni scholarship from the Juntendo University School of Medicine, research

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fellowship from Uehara Memorial Foundation, Pelotonia postdoctoral fellowship (to TS), and

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the Blue Beautiful Skies Fund (to DPC).

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References 1. Foundation Medicine. http://www.foundationmedicine.com/. last accessed on 27/Jun/2017.

non-small cell lung cancer. Fam Cancer. 2015 Sep; 14(3): 463-9.

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2. Clamon GH, Bossler AD, Abu Hejleh T, et al. Germline mutations predisposing to

3. Guardant Health. https://www.guardanthealth.com/guardant360/. last accessed on

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27/Jun/2017.

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Figure legends Figure 1. Pathological features of small cell lung carcinoma in a 50-year-old female never

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smoker, left upper lobe wedge resection; hematoxylin-eosin (A), immunohistochemistry for

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synaptophysin (B) and thyroid transcription factor-1 (C); original magnification x400.

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Table 1 Results of FoundationOneTM in a 50-year-old female never smoker with recurrent small-cell lung cancer Lung Cancer Specimen (collected and submitted in 2015) Alterations

Mutation allele frequency 83%

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TP53 Y236* PARK2 Q347*

47%

FGFR2 Amplification

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Breast Cancer Specimen (collected in 2010, submitted in 2015) Alterations

Mutation allele frequency

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TP53 Y236* PARK2 Q347* STK11 truncation exon8 CCND3 amplification

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VEGFA amplification

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Abbreviation: NA, not available.

NA NA NA NA NA

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Table 2 Result of FoundationACTTM in a 34-year-old female former smoker with advanced lung adenocarcinoma Blood (collected and submitted in 2016) Alterations

Mutation allele frequency

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MET 3028+2T>C (splice site mutation)

0.19%

50.7%

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BRCA2 L3061*

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