From epithelial remodelling to carcinogenesis

Journal Pre-proof From epithelial remodelling to carcinogenesis

Leszek Satora, Jennifer Mytych, Anna Bilska-Kos, Katarzyna Kozioł PII:

S0079-6107(19)30080-X

DOI:

https://doi.org/10.1016/j.pbiomolbio.2019.08.001

Reference:

JPBM 1469

To appear in:

Progress in Biophysics and Molecular Biology

Received Date:

16 April 2019

Accepted Date:

01 August 2019

Please cite this article as: Leszek Satora, Jennifer Mytych, Anna Bilska-Kos, Katarzyna Kozioł, From epithelial remodelling to carcinogenesis, Progress in Biophysics and Molecular Biology (2019), https://doi.org/10.1016/j.pbiomolbio.2019.08.001

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From epithelial remodelling to carcinogenesis

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*Leszek

Satoraa, Jennifer Mytychb, Anna Bilska-Kosc and Katarzyna Koziołb

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*aPomeranian

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bDepartment

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36-100 Kolbuszowa, Poland.

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cDepartment

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Institute - National Research Institute, Radzików, 05-870 Błonie, Poland

Center of Clinical Toxicology Kartuska 4/6, 80-104 Gdansk, Poland.

of Animal Physiology and Reproduction, University of Rzeszów, Werynia 502,

of Plant Biochemistry and Physiology, Plant Breeding and Acclimatization

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*Corresponding

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Dr Leszek Satora

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Pomeranian Center of Clinical Toxicology Kartuska 4/6, 80-104 Gdansk, Poland.

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Tel: +48 682 19 39

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E-mail: [email protected]

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author

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Abstract

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The novel cancer theory named ‘The tissue organization field theory’ (TOFT) suggests that

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carcinogenesis is a process analogous to embryonic development, whereby organs are formed

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through interactions among different cell types. The suggested ‘morphological remodelling’

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of the epithelium under hypoxia in gut breathing fish (GBF) has many common features with

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carcinogenesis. It appears that research into the relationship among epidermal growth factor

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receptor (EGFR), hypoxia inducible factor (HIF) as well as hypoxia and normoxia states in

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GBF fishes can be crucial in learning about the steering mechanisms of squamous epithelium

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proliferation, leading to a better understanding of carcinogenesis.

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Key Words: Gut breathing fish; EGFR, HIF 1; hypoxia; epithelial remodelling;

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carcinogenesis; TOFT theory

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The proliferation of the squamous cells in the digestive tract of gut breathing fish (GBF)

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under hypoxia conditions seems to be an extremely beneficial event. This process allows fish

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to create a thin air-blood barrier and use the digestive tract as an additional respiratory organ

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during hypoxic conditions (Satora et al., 2019). Ancistrus chagresi (spinosus) included in the

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GBF fish group can adjust about 25% of the surface of the stomach to air-breathing (Graham,

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1997), while in bronze Corydoras about 48% of the digestive tract can be transformed into an

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additional respiratory organ under water hypoxia conditions (Leknes, 2015). Therefore it may

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be indicated that the above mentioned adaptations are a type of the‘morphological

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remodelling’ of epithelial cells.

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Studies using specific antibodies against the intracellular domain of human epidermal growth

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factor receptor (EGFR) demonstrated the presence of active, proliferating squamous cells in

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the GBF fish digestive tract, in both corydoras (Satora et al., 2017; Mytych et al., 2018) and

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otocinclus (Satora et al., 2019). High EGFR expression observed in the respiratory part of the

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intestine in bronze Corydoras and in the stomach of O. affinis suggest higher proliferation

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activity and angiogenesis of epithelium in that part of the intestine, which creates the

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conditions for aerial respiration. Furthermore, quantitative analysis of EGFR protein

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abundance also revealed its higher synthesis in the respiratory stomach of O. affinis (Satora et

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al., 2019). However, the experimental setup allowing inhibition and /or stimulation of

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proliferation markers seems to be an important next stage of research on gut breathing fish.

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Antibodies anti-EGFR (cytoplasmic domain) are used to test active, proliferating epithelial

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cells in the diagnosis, prognosis and monitoring of the squamous carcinomas (Gröbe et al.,

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2014). Increased levels of EGFR are linked with malignant transformations of squamous cells

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(e.g. squamous cell carcinoma in the lungs, esophagus, neck, head). EGFR mediating

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signalling has been shown to induce mitogenic responses in a variety of cell types. The

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diagnosis of poor prognosis is based on the studies of EGFR expression in human breast

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cancer (Gröbe et al., 2014). Most studies on EGFR and its ligands in fish were performed

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using recombinant mammalian proteins (Wang and Ge, 2004; Tse and Ge, 2009; Mytych et

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al., 2018). Furthermore, in teleost fish the lineage-specific co-evolution of the EGF

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receptor/ligand signalling system was demonstrated (Laisney et al., 2010) and the

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amplification of EGFR gene was also observed (Gomez et al., 2004; Laisney et al., 2010),

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similarly to carcinogenesis (Chang et al., 2019). It may be speculated that this process is

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induced by hypoxia.

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Hypoxia inducible factor-1 (HIF-1) is one of the major components of cellular response to

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oxygen deprivation. Activated HIF-1 plays a crucial role in adaptive responses of the tumour

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cells to fluctuations in oxygen concentration through transcriptional activation of over 100

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downstream genes. HIF-1 regulates important biological processes required for tumour

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survival and progression, including transcription of genes involved in the cell proliferation

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(Masoud and Li, 2015).

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The PCR studies and immunolocalization using antibodies specific to HIF-1α followed by

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visualization under transmission electron microscope (TEM) revealed high expression of HIF-

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1α and the presence of numerous HIF-1α epitopes in the respiratory part of the bronze

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Corydoras’ intestine (Satora et al., 2018). In all GBF fish species examined a potential ability

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for proliferation of squamous cells in various parts of the intestine was observed. It seems that

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it is carried out according to the same mechanism, being driven by the states of water hypoxia

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inducing a ‘cascade of events’ which leads to the proliferation of squamous epithelial cells

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(Satora et al., 2017; Mytych et al., 2018; Satora et al., 2018; Satora et al., 2019).

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The appearance of tetrapods and land colonization, one of the most important occurrence in

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vertebrate evolution, were both connected with the high capacity of the air-breathing

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(Graham, 1997). Although there is no direct physiological evidence described in the

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paleontological records and we can only study currently existing species, a number of

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generalizations can be made. For example, adapting the digestive tract to function as an

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additional respiratory organ under hypoxia states seems to be associated with numerous

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difficulties in the development of this ability in GBF fishes (Nelson, 2014). The extremely

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delicate respiratory epithelium; the minimum thickness of the air-blood barrier in GBF fish is

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about 0.2 m; is highly susceptible to mechanical damage and the action of gastric secretions

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(Satora, 1998; Nelson, 2014). On the other hand, the ability to proliferate squamous cells and

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the ‘morphological remodelling’ of the epithelium of the digestive tract in low oxygen

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conditions seems to be an excellent property. Phylogenetically, the lungs were derived as a

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unique adaptation to conditions of low oxygen content in the aquatic medium wherein the fish

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ancestors lived. Despite differences, all of these organisms possessed a characteristic feature:

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the presence of a simple squamous epithelium covering the numerous capillary vessels. This

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adaptation facilitates gas diffusion (Satora, 1998; Icardo, 2018).

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Lungs as well as respiratory and non-respiratory bladders of chondrostean appear to originate

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from a respiratory, posterior pharynx (Icardo, 2018). It can also be assumed that their

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formation could be related to the proliferation of the squamous cells and the gradual

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enlargement of the diverticulum in the anterior part of the digestive tract. If the proposed

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‘morphological remodelling’ process, which involves the proliferation of squamous cells

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under hypoxia, played an important role in the development of lung and gas bladders, it could

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have been further consolidated. It is suggested, that in GBF fishes the proliferation process of

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squamous cells is strictly controlled and is probably inhibited under normoxia. It also seems

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that putative O2 chemoreceptors, the neuroepithelial cells (NECs) which manifest peculiar

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neurotransmitter profiles, they can play an important role in this process. However, future

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experiments are needed to evaluate how NECs and neural pathways are centrally integrated in

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GBF fishes (Zaccone et al., 2006; Jonz, 2018; Satora et al., 2019). In mammals during lung

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development, undifferentiated cuboidal cells form lining in distal air spaces and transform

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into respiratory epithelium (pneumocytes). EGF receptors are of critical importance in this

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process (Miettinen et al., 1997).

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About 90 years ago Otto Warburg described how cancer cells avidly consume glucose and

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produce lactic acid under aerobic conditions (Warburg effect). Hypoxia, frequently observed

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in solid tumors, results in an increased HIF1 transcription factor activity. Furthermore, HIF-

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1is one of the most important effector molecules downstream to EGFR pathways (Koppenol

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et al., 2011). On the other hand, hypoxia and normoxia are opposed conditions adjusting gill

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remodelling as well as the production or reduction of cellular mass between the lamellae of

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the gills in several species of teleosts fish (Nilsson, 2007). In this process NECs – putative O2

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chemoreceptors - are involved (Tzaneva et al., 2001).

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In accordance with the novel theory, cancer is a relational problem. The main attention was

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focused on the level of biological organization - cancer as a problem of tissue organization

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similar to histogenesis and organogenesis (Sonnenschein and Soto, 2016). According to

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Sonnenschein and Soto (2016), carcinogenesis is a process analogous to embryonic

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development, whereby organs are formed through interactions of different cell types. The

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basic premise of the tissue organization field theory (TOFT) is that proliferation with

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variation and motility is the default state of all cells (Sonnenschein and Soto, 2016).

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On the other hand, epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial

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transition (MET) processes, take part not only in the physiological conditions but also in the

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carcinogenesis (Taylor et al., 2010; Chaffer et al., 2007; Hugo et al., 2007). Further, it has

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been demonstrated, that EMT may be induced by hypoxia states and through activation of the

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EGFR signalling pathway (Misra et al., 2012). In addition, HIFs and EGFR are overexpressed

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in many types of cancer (Wang 2017; Talks et al., 2000; Zhong et al.,1999) and exhibit, i.a.

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mitogenic effects (Hirsch et al.,2003; Schultz et al., 2006; Gordan et al., 2007). There is a

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possibility that enterocytes in the GBF fishes undergo a process of EMT under hypoxia. In

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this scenario, during EMT process, the cells lose intercellular connection and acquire ability

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to move, then they transform into mesenchymal cells with increased proliferation ability, and

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next, they may differentiate into various cell types (including squamous) during MET process

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(Kalluri and Weinberg, 2009; Pei et al., 2019). The resulting squamous cells are the optimal

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barrier for gas exchange in the gastrointestinal tract. EMT and/or MET are accompanied by

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serial events of extracellular matrix deposition, changes in the expression of genes associated

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with both epithelium or mesenchymal layers, and eventually changes in the morphology

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(Moustakas and Heldin, 2007). It is crucial to maintain the balance between proliferative and

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antiproliferative factors and the correct regulation of cellular plasticity through EMT and

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MET under local cellular environment (Gérard and Goldbeter, 2014; Roca et al., 2013). It is

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suggested that both EGFR and HIF-1α may be essential signals enabling the ‘default state’ of

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gastrointestinal epithelial cells in GBF fishes, in the response to the change in the oxygen

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concentrations. Finally, there is the morphological rebuilding of the epithelium, which seems

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to be consistent with the TOFT theory.

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The suggested ‘morphological remodelling’ of the epithelium under hypoxia – factor which

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‘triggers’ cells from the state of equilibriumin gut breathing fish has a number of common

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features with carcinogenesis. However, it should be emphasized that the mechanisms of

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dependence of metabolic profiles and cellular phenotypes are still poorly recognized as well

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as the regulation of the fate of the cells in the cell proliferation processes is only just

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beginning to be understood. Thus, GBF fishes are not only an evolutionary curiosity that

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comes as a surprise to many researches (Icardo, 2018), but they also seem to be a very

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interesting model organism within which the proliferation of squamous cells can be examined

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in a multithreaded way. Further investigations of the mechanisms controlling this

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phenomenon in gut breathing fish will result in extending our knowledge about cancer

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processes.

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Conflict of interest

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The authors declare no conflict of interest.

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Author contributions

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L.S. conceived and designed the research project and wrote the manuscript. J.M., A.B-K and

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K.K. interpreted the results and helped writing the manuscript.

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Journal Pre-proof Dr Leszek Satora Kraków 15.04.2019 Pomeranian Center of Clinical Toxicology Kartuska 4/6, 80-104 Gdansk, Poland. The Editor Progress in Biophysics & Molecular Biology The manuscript entitled “From epithelial remodeling to carcinogenesis” has not any conflicts of interest.

Sincerely, On behalf of all authors, Leszek Satora

Journal Pre-proof 

The renowned biochemist Otto Warburg described how cancer cells avidly consume glucose and produce lactic acid under aerobic conditions (Warburg effect).



Hypoxia, frequently observed in solid tumors, results in an increased HIF1 transcription factor activity. Furthermore, HIF-1is one of the most important effector molecules downstream to EGFR pathways.



The suggested “morphological remodelling” of the epithelium under hypoxia in gut breathing fish (GBF) has a number of common features with carcinogenesis.



The observed similarities in the proliferation of squamous cells in GBF seem to confirm the ‘tissue organization field theory’ proposed by Sonnenschein and Soto.



Gut breathing fish seem to be a very interesting model organism within which the proliferation of squamous cells can be examined in a multithreaded way.



Further investigations of the mechanisms controlling this phenomenon in GBF will result in a better understanding of cancer processes.