Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Sep;258(1):69-82.
doi: 10.1002/path.5978. Epub 2022 Jul 18.

Peak density of immature nerve cells occurs with high-grade dysplasia in intraductal papillary mucinous neoplasms of the pancreas

Vincent Quoc-Huy Trinh  1 , Joseph Thomas Roland  1   2 , Jahg Wong  3 , Frank Revetta  4 , Krutika Patel  4 , Chanjuan Shi  5 , Kathleen E DelGiorno  2   6   7 , Bruce D Carter  8 , Marcus Chuan Beng Tan  1   7   9
Affiliations

Peak density of immature nerve cells occurs with high-grade dysplasia in intraductal papillary mucinous neoplasms of the pancreas

Vincent Quoc-Huy Trinh et al. J Pathol. 2022 Sep.

Abstract

The development of neural structures within tumors is now considered vital for carcinogenesis. However, the time course of this development in human pre-invasive neoplasia has been incompletely described. Therefore, we performed a detailed analysis of nerves across the neoplastic spectrum in resected intraductal papillary mucinous neoplasms (IPMNs) of the pancreas. Histology and multiplexed immunochemistry demonstrated that nerve density increased from low-grade (LG) to high-grade dysplasia (HG) but did not further increase once invasive IPMN (INV IPMN) was present. Higher nerve density correlated with increasing expression of nerve growth factor (NGF) by the tumor cells. Intra-tumoral nerves were immature and lacked markers of sympathetic, parasympathetic, and sensory lineages. Here, we show for the first time the presence of neural precursor cells (NPCs) within the stroma of pancreatic tumors. The density of these doublecortin (DCX)-positive NPCs increased from LG to HG, but not from HG to INV IPMN. We conclude that peak neural density of tumors is reached in high-grade dysplasia (often termed carcinoma in situ) rather than after invasion. These findings suggest that nerve-tumor interactions are important in IPMN progression and may serve as the basis for future mechanistic studies and novel therapeutic modalities. © 2022 The Pathological Society of Great Britain and Ireland.

Keywords: intraductal papillary mucinous neoplasms; neoplasms; nerves; neural factors; pancreas.

PubMed Disclaimer

Conflict of interest statement

No conflicts of interest were declared

Figures

Figure 1.
Figure 1.. Increased stromal innervation with HG IPMNs.
(A) IPMN stromal innervation as graded by H&E interpretation by two pathologists. Scale bar, 200 μm; dashed 200 × 200 μm squares are magnified on the right with dashed outline representing nerves. G1: rare, isolated nerve bundles (arrowhead); G2: increased frequency of nerve bundles with moderate increase in size; G3: overt hyperplasia of nerves in the IPMN stroma with enlarged and hyperplastic nerves (star). (B) Stromal innervation according to IPMN grade. LG versus HG p=0.0012; HG versus INV IPMN p=0.493; Kruskal–Wallis (thick straight line) p=0.0026. (C) Vimentin immunohistochemistry-based mapping of stroma, epithelium, parenchyma and “glass” generated by machine learning in Ilastik and Cellprofiler. Scale bar on the left image, 2 mm; on the right image, 100 μm. (D) Left: Examples of double-positive S100 and GAP-43 structures larger than 500 pixels (23.53 μm2). Scale bar, 200 μm. Right: Examples of double positive S100 and GAP-43 double-positive small structures (less than 500 pixels, 23.53 μm2); scale bar, 50 μm. (E) Number of large S100+ GAP-43+ according to IPMN grade, LG versus HG p=0.0010, LG versus INV IPMN p=0.0456. (F) There were more small S100+ GAP-43+ structures in HG IPMN compared to LG (p=0.0222). Abbreviations: G1–3: Grades 1–3. NP: normal parenchyma. LG: low-grade IPMN. HG: high-grade IPMN. INV IPMN: invasive IPMN.
Figure 2.
Figure 2.. NGF expression in IPMN correlated with grade of dysplasia, and NGF receptors were expressed by nerves in the tumor stroma.
(A) Tumor cell NGF expression according to IPMN grade, Kruskal–Wallis (thick straight line) p<0.0001, LG versus HG p=0.0012, HG versus INV IPMN p=0.0055. (B) TrkA expression in nerves inside and outside the IPMN stroma, Kruskal–Wallis (thick straight line) p=0.0921. (C) p75NTR expression, Kruskal–Wallis p<0.0001, LG adjacent versus stroma p<0.0001, HG adjacent versus stroma p<0.0001, INV IPMN adjacent versus stroma p=0.0008. Image squares measure 200 μm × 200 μm. Abbreviations: NP: normal parenchyma duct, LG: low-grade IPMN, HG: high-grade IPMN, INV IPMN: invasive IPMN.
Figure 3.
Figure 3.. Nerves inside the IPMN stroma express markers differently from nerves located away from the IPMN.
Low-power view of tyrosine hydroxylase staining in a low-grade intestinal IPMN section; scale bar, 2 mm. The black arrowheads identify nerve bundles located within the stroma of IPMNs, while the red arrow heads identify nerves in the adjacent normal pancreatic parenchyma. Black and red 200 × 200 μm squares are magnified in bottom-left (intra-tumoral stroma) and bottom-right images (adjacent tissue parenchyma), with the nerve highlighted by the dashed line.
Figure 4.
Figure 4.. Nerve signaling according to their location inside and outside the IPMN stroma.
(A) TH (catecholamine-driven nerve marker) expression in nerves, Kruskal–Wallis p=0.0020, LG adjacent versus stroma p=0.0095, HG adjacent versus stroma p=0.0118, INV IPMN adjacent versus stroma p=0.8181. (B) VAchT (cholinergic-driven nerve marker) expression in nerves, Kruskal–Wallis p=0.9998. (C) CGRP (sensory nerve marker) expression in nerves, Kruskal–Wallis p=0.3350. (D) Nestin (marker of immature nerves) expression, Kruskal–Wallis p<0.0001, LG adjacent versus stroma p<0.0001, HG adjacent versus stroma p=0.0030, INV IPMN adjacent versus stroma p=0.0003; LG stroma versus HG stroma p=0.4969, LG stroma versus INV IPMN stroma p=0.0100. All image squares measure 200 μm × 200 μm. Abbreviations: TH: Tyrosine hydroxylase, CGRP: calcitonin gene-related peptide, LG: low-grade IPMN, HG: high-grade IPMN, INV IPMN: invasive IPMN.
Figure 5.
Figure 5.. PDAC stromal nerves have stronger TH expression compared to IPMN.
(A) Examples of invaded nerves located in invasive IPMN and in PDAC samples; scale bar 40 μm. (B) Stromal TH in nerves, p=0.0275. (C) Stromal VAchT in nerves, p=1.000. (D) Stromal CGRP in nerves, p=0.4067. (E) Stromal Nestin in nerves, p=1.000. Abbreviations: TH: Tyrosine hydroxylase, CGRP: calcitonin gene-related peptide, LG: low-grade IPMN, HG: high-grade IPMN, INV IPMN: invasive IPMN.
Figure 6.
Figure 6.. DCX-positive stromal cells have increased density in high-grade and invasive IPMNs.
(A) Example of DCX-positive cells in normal parenchyma (NP, left) and underlying a HG-IPMN (right). B) Count of DCX+ large stromal cells in ten 20X objective fields (68,000 μm2) of stroma, from 6 periductular areas around normal parenchyma, 11 LG-IPMN, 11 HG-IPMN, and 7 invasive IPMN. NP versus LG p<0.0001; LG versus HG p=0.0031; NP versus HG p<0.0001; HG versus INV IPMN p=0.1120. C) Co-expression of DCX with PSA-NCAM was noted in large stromal cells; scale bar, 100 μm).
See this image and copyright information in PMC

References

    1. Maitra A, Fukushima N, Takaori K, et al. Precursors to invasive pancreatic cancer. Adv Anat Pathol 2005; 12: 81–91 - PubMed
    1. Muraki T, Jang K-T, Reid MD, et al. Pancreatic ductal adenocarcinomas associated with intraductal papillary mucinous neoplasms (IPMNs) versus pseudo-IPMNs: relative frequency, clinicopathologic characteristics and differential diagnosis. Mod Pathol September 2021 - PubMed
    1. Yopp AC, Katabi N, Janakos M, et al. Invasive Carcinoma Arising in Intraductal Papillary Mucinous Neoplasms of the Pancreas: A Matched Control Study with Conventional Pancreatic Ductal Adenocarcinoma. Ann Surg 2011; 253: 968–974 - PubMed
    1. Salvia R, Burelli A, Perri G, et al. State-of-the-art surgical treatment of IPMNs. Langenbecks Arch Surg November 2021 - PMC - PubMed
    1. Tanaka M International consensus on the management of intraductal papillary mucinous neoplasm of the pancreas. Ann Transl Med 2015; 3: 9. - PMC - PubMed

Publication types

MeSH terms