Spatial profiling of gastric cancer patient-matched primary and locoregional metastases reveals principles of tumour dissemination

R. Sundar; D.H.W. Liu; G.G.A. Hutchins; H.L. Slaney; A.N.S. Silva; J. Oosting; J.D. Hayden; L.C. Hewitt; C.C.Y. Ng; A. Mangalvedhekar; S.B. Ng; I.B.H. Tan; P. Tan; H.I. Grabsch

doi:10.1136/gutjnl-2020-320805

Spatial profiling of gastric cancer patient-matched primary and locoregional metastases reveals principles of tumour dissemination

R. Sundar, D.H.W. Liu, G.G.A. Hutchins, H.L. Slaney, A.N.S. Silva, J. Oosting, J.D. Hayden, L.C. Hewitt, C.C.Y. Ng, A. Mangalvedhekar, S.B. Ng, I.B.H. Tan^*, P. Tan^*, H.I. Grabsch^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Objective Endoscopic mucosal biopsies of primary gastric cancers (GCs) are used to guide diagnosis, biomarker testing and treatment. Spatial intratumoural heterogeneity (ITH) may influence biopsy-derived information. We aimed to study ITH of primary GCs and matched lymph node metastasis (LNmet).Design GC resection samples were annotated to identify primary tumour superficial (PTsup), primary tumour deep (PTdeep) and LN met subregions. For each subregion, we determined (1) transcriptomic profiles (NanoString 'PanCancer Progression Panel', 770 genes); (2) next-generation sequencing (NGS, 225 gastrointestinal cancer-related genes); (3) DNA copy number profiles by multiplex ligation-dependent probe amplification (MLPA, 16 genes); and (4) histomorphological phenotypes.Results NanoString profiling of 64 GCs revealed no differences between PT sup1 and PT (sup2), while 43% of genes were differentially expressed between PT sup versus PTdeep and 38% in PT (sup) versus LNmet. Only 16% of genes were differently expressed between PTdeep and LNmet. Several genes with therapeutic potential (eg IGF1, PIK3CD and TGFB1) were overexpressed in LNmet and PTdeep compared with PT sup. NGS data revealed orthogonal support of NanoString results with 40% mutations present in PT deep and/or LNmet, but not in PT (sup). Conversely, only 6% of mutations were present in PT sup and were absent in PTdeep and LNmet. MLPA demonstrated significant ITH between subregions and progressive genomic changes from PTsup to PTdeep/LN met.Conclusion In GC, regional lymph node metastases are likely to originate from deeper subregions of the primary tumour. Future clinical trials of novel targeted therapies must consider assessment of deeper subregions of the primary tumour and/or metastases as several therapeutically relevant genes are only mutated, overexpressed or amplified in these regions.

Original language	English
Pages (from-to)	1823-1832
Number of pages	10
Journal	Gut
Volume	70
Issue number	10
DOIs	https://doi.org/10.1136/gutjnl-2020-320805
Publication status	Published - 1 Oct 2021

Keywords

GENOMIC HETEROGENEITY
GENETIC PROFILES
LYMPH-NODES

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Sundar, R., Liu, D. H. W., Hutchins, G. G. A., Slaney, H. L., Silva, A. N. S., Oosting, J., Hayden, J. D., Hewitt, L. C., Ng, C. C. Y., Mangalvedhekar, A., Ng, S. B., Tan, I. B. H., Tan, P., & Grabsch, H. I. (2021). Spatial profiling of gastric cancer patient-matched primary and locoregional metastases reveals principles of tumour dissemination. Gut, 70(10), 1823-1832. https://doi.org/10.1136/gutjnl-2020-320805

@article{d4d204aa5de940a39b010e66934bf979,

title = "Spatial profiling of gastric cancer patient-matched primary and locoregional metastases reveals principles of tumour dissemination",

abstract = "Objective Endoscopic mucosal biopsies of primary gastric cancers (GCs) are used to guide diagnosis, biomarker testing and treatment. Spatial intratumoural heterogeneity (ITH) may influence biopsy-derived information. We aimed to study ITH of primary GCs and matched lymph node metastasis (LNmet).Design GC resection samples were annotated to identify primary tumour superficial (PTsup), primary tumour deep (PTdeep) and LN met subregions. For each subregion, we determined (1) transcriptomic profiles (NanoString 'PanCancer Progression Panel', 770 genes); (2) next-generation sequencing (NGS, 225 gastrointestinal cancer-related genes); (3) DNA copy number profiles by multiplex ligation-dependent probe amplification (MLPA, 16 genes); and (4) histomorphological phenotypes.Results NanoString profiling of 64 GCs revealed no differences between PT sup1 and PT (sup2), while 43% of genes were differentially expressed between PT sup versus PTdeep and 38% in PT (sup) versus LNmet. Only 16% of genes were differently expressed between PTdeep and LNmet. Several genes with therapeutic potential (eg IGF1, PIK3CD and TGFB1) were overexpressed in LNmet and PTdeep compared with PT sup. NGS data revealed orthogonal support of NanoString results with 40% mutations present in PT deep and/or LNmet, but not in PT (sup). Conversely, only 6% of mutations were present in PT sup and were absent in PTdeep and LNmet. MLPA demonstrated significant ITH between subregions and progressive genomic changes from PTsup to PTdeep/LN met.Conclusion In GC, regional lymph node metastases are likely to originate from deeper subregions of the primary tumour. Future clinical trials of novel targeted therapies must consider assessment of deeper subregions of the primary tumour and/or metastases as several therapeutically relevant genes are only mutated, overexpressed or amplified in these regions.",

keywords = "GENOMIC HETEROGENEITY, GENETIC PROFILES, LYMPH-NODES",

author = "R. Sundar and D.H.W. Liu and G.G.A. Hutchins and H.L. Slaney and A.N.S. Silva and J. Oosting and J.D. Hayden and L.C. Hewitt and C.C.Y. Ng and A. Mangalvedhekar and S.B. Ng and I.B.H. Tan and P. Tan and H.I. Grabsch",

note = "Funding Information: Funding RS is supported by a National Medical Research Council (NMRC) Fellowship, Singapore. PT is supported by Duke-NUS Medical School and the Genome Institute of Singapore, Agency for Science, Technology and Research. This work was also supported by National Medical Research Council grants (OF-LCG18May-0023, NR13NMR111OM and NMRC/STaR/0026/2015). This study was supported by Pathological Society Career Development Fellowship (CDF2015/01, PI: GH) and Academy of Medical Sciences Starter Grant for Clinical Lecturers (SGCL14/ GH, PI: GH). Funding Information: Competing interests PT has stock and other ownership interests in Tempus Healthcare, research funding from Kyowa Hakko Kirin and Thermo Fisher Scientific, and patents/other intellectual property through the Agency for Science and Technology Research, Singapore (all outside the submitted work). RS has received honoraria from Bristol-Myers Squibb, Lilly and MSD, has advisory activity with Bristol-Myers Squibb, Eisai and AstraZeneca, received research funding from Paxman Coolers and has received travel grants from AstraZeneca, Roche and Taiho Pharmaceutical (all outside the submitted work). IBHT has received consultation fees for MSD, Taiho, Roche, BMS, Bayer, Amgen and Merck Serono, and research funding from MSD and Taiho (all outside the submitted work). HIG has received honoraria from MSD (outside of the submitted work). Patient consent for publication Not required. Ethics approval The study was approved by the Leeds Research Ethics Committee (LREC no. CA01/122). Provenance and peer review Not commissioned; externally peer reviewed. Publisher Copyright: {\textcopyright} ",

year = "2021",

month = oct,

day = "1",

doi = "10.1136/gutjnl-2020-320805",

language = "English",

volume = "70",

pages = "1823--1832",

journal = "Gut",

issn = "0017-5749",

publisher = "BMJ Publishing Group",

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Sundar, R, Liu, DHW, Hutchins, GGA, Slaney, HL, Silva, ANS, Oosting, J, Hayden, JD, Hewitt, LC, Ng, CCY, Mangalvedhekar, A, Ng, SB, Tan, IBH, Tan, P & Grabsch, HI 2021, 'Spatial profiling of gastric cancer patient-matched primary and locoregional metastases reveals principles of tumour dissemination', Gut, vol. 70, no. 10, pp. 1823-1832. https://doi.org/10.1136/gutjnl-2020-320805

TY - JOUR

T1 - Spatial profiling of gastric cancer patient-matched primary and locoregional metastases reveals principles of tumour dissemination

AU - Sundar, R.

AU - Liu, D.H.W.

AU - Hutchins, G.G.A.

AU - Slaney, H.L.

AU - Silva, A.N.S.

AU - Oosting, J.

AU - Hayden, J.D.

AU - Hewitt, L.C.

AU - Ng, C.C.Y.

AU - Mangalvedhekar, A.

AU - Ng, S.B.

AU - Tan, I.B.H.

AU - Tan, P.

AU - Grabsch, H.I.

N1 - Funding Information: Funding RS is supported by a National Medical Research Council (NMRC) Fellowship, Singapore. PT is supported by Duke-NUS Medical School and the Genome Institute of Singapore, Agency for Science, Technology and Research. This work was also supported by National Medical Research Council grants (OF-LCG18May-0023, NR13NMR111OM and NMRC/STaR/0026/2015). This study was supported by Pathological Society Career Development Fellowship (CDF2015/01, PI: GH) and Academy of Medical Sciences Starter Grant for Clinical Lecturers (SGCL14/ GH, PI: GH). Funding Information: Competing interests PT has stock and other ownership interests in Tempus Healthcare, research funding from Kyowa Hakko Kirin and Thermo Fisher Scientific, and patents/other intellectual property through the Agency for Science and Technology Research, Singapore (all outside the submitted work). RS has received honoraria from Bristol-Myers Squibb, Lilly and MSD, has advisory activity with Bristol-Myers Squibb, Eisai and AstraZeneca, received research funding from Paxman Coolers and has received travel grants from AstraZeneca, Roche and Taiho Pharmaceutical (all outside the submitted work). IBHT has received consultation fees for MSD, Taiho, Roche, BMS, Bayer, Amgen and Merck Serono, and research funding from MSD and Taiho (all outside the submitted work). HIG has received honoraria from MSD (outside of the submitted work). Patient consent for publication Not required. Ethics approval The study was approved by the Leeds Research Ethics Committee (LREC no. CA01/122). Provenance and peer review Not commissioned; externally peer reviewed. Publisher Copyright: ©

PY - 2021/10/1

Y1 - 2021/10/1

N2 - Objective Endoscopic mucosal biopsies of primary gastric cancers (GCs) are used to guide diagnosis, biomarker testing and treatment. Spatial intratumoural heterogeneity (ITH) may influence biopsy-derived information. We aimed to study ITH of primary GCs and matched lymph node metastasis (LNmet).Design GC resection samples were annotated to identify primary tumour superficial (PTsup), primary tumour deep (PTdeep) and LN met subregions. For each subregion, we determined (1) transcriptomic profiles (NanoString 'PanCancer Progression Panel', 770 genes); (2) next-generation sequencing (NGS, 225 gastrointestinal cancer-related genes); (3) DNA copy number profiles by multiplex ligation-dependent probe amplification (MLPA, 16 genes); and (4) histomorphological phenotypes.Results NanoString profiling of 64 GCs revealed no differences between PT sup1 and PT (sup2), while 43% of genes were differentially expressed between PT sup versus PTdeep and 38% in PT (sup) versus LNmet. Only 16% of genes were differently expressed between PTdeep and LNmet. Several genes with therapeutic potential (eg IGF1, PIK3CD and TGFB1) were overexpressed in LNmet and PTdeep compared with PT sup. NGS data revealed orthogonal support of NanoString results with 40% mutations present in PT deep and/or LNmet, but not in PT (sup). Conversely, only 6% of mutations were present in PT sup and were absent in PTdeep and LNmet. MLPA demonstrated significant ITH between subregions and progressive genomic changes from PTsup to PTdeep/LN met.Conclusion In GC, regional lymph node metastases are likely to originate from deeper subregions of the primary tumour. Future clinical trials of novel targeted therapies must consider assessment of deeper subregions of the primary tumour and/or metastases as several therapeutically relevant genes are only mutated, overexpressed or amplified in these regions.

AB - Objective Endoscopic mucosal biopsies of primary gastric cancers (GCs) are used to guide diagnosis, biomarker testing and treatment. Spatial intratumoural heterogeneity (ITH) may influence biopsy-derived information. We aimed to study ITH of primary GCs and matched lymph node metastasis (LNmet).Design GC resection samples were annotated to identify primary tumour superficial (PTsup), primary tumour deep (PTdeep) and LN met subregions. For each subregion, we determined (1) transcriptomic profiles (NanoString 'PanCancer Progression Panel', 770 genes); (2) next-generation sequencing (NGS, 225 gastrointestinal cancer-related genes); (3) DNA copy number profiles by multiplex ligation-dependent probe amplification (MLPA, 16 genes); and (4) histomorphological phenotypes.Results NanoString profiling of 64 GCs revealed no differences between PT sup1 and PT (sup2), while 43% of genes were differentially expressed between PT sup versus PTdeep and 38% in PT (sup) versus LNmet. Only 16% of genes were differently expressed between PTdeep and LNmet. Several genes with therapeutic potential (eg IGF1, PIK3CD and TGFB1) were overexpressed in LNmet and PTdeep compared with PT sup. NGS data revealed orthogonal support of NanoString results with 40% mutations present in PT deep and/or LNmet, but not in PT (sup). Conversely, only 6% of mutations were present in PT sup and were absent in PTdeep and LNmet. MLPA demonstrated significant ITH between subregions and progressive genomic changes from PTsup to PTdeep/LN met.Conclusion In GC, regional lymph node metastases are likely to originate from deeper subregions of the primary tumour. Future clinical trials of novel targeted therapies must consider assessment of deeper subregions of the primary tumour and/or metastases as several therapeutically relevant genes are only mutated, overexpressed or amplified in these regions.

KW - GENOMIC HETEROGENEITY

KW - GENETIC PROFILES

KW - LYMPH-NODES

U2 - 10.1136/gutjnl-2020-320805

DO - 10.1136/gutjnl-2020-320805

M3 - Article

C2 - 33229445

SN - 0017-5749

VL - 70

SP - 1823

EP - 1832

JO - Gut

JF - Gut

IS - 10

ER -