By mimicking in-vivo environments, 3D cell culture models appear nowadays as the best in-vitro study model to work in an in-vivo like study model and to obtain more physiologically relevant data and proof of concept as close as possible to “a clinical context” (aka the “near-human” approach).

3D cell culture chip – AIM Biotech tebu-bio – near-human approaches

To accompany researchers along this way, there are several innovative cell culture devices (available in Europe through tebu-bio), and more specifically a modular microfluidic platform for 3D cell culture with the capability to monitor complex biological systems dynamics in response to tuned microenvironment: the 3D Cell culture chip (Aim Biotech)

Classically, it was known that CD4⁺ T cells, upon activation and expansion, develop into different T helper cell subsets with different cytokine profiles and distinct effector functions to effect the immune response. Until some years ago, T cells were divided into Th1 or Th2 cells, depending on the cytokines they produce (1).

However, a third subset of IL-17-producing effector T helper cells, called Th17 cells, was discovered and characterised. Th17 cells produce several factors that can be quantified with one single multiplex immunoassay (e.g. IL-17, IL-17F, and IL-22). Th17 cells seem to induce a massive tissue reaction owing to the broad distribution of the IL-17 and IL-22 receptors.

Th17 cells also secrete IL-21 to communicate with the cells of the immune system. The differentiation factors (TGF-β plus IL-6 or IL-21), the growth and stabilization factor (IL-23), and the transcription factors (STAT3, RORγt, and RORα) involved in the development of Th17 cells have also been identified. Th17 cells thereby seem to be mainly involved in clearing pathogens during host defense reactions and in inducing tissue inflammation in autoimmune disease.

Let’s focus on the role of TGF-β. Though it has been shown to positively regulate the development of murine T helper type 17 (Th17) cells, which of the intracellular signaling pathways are involved is controversial. A recent publication using protein antibody arrays found an increase of expression and phosphorylation of the following Smad-independent signaling molecules in Th17-polarized wild-type T cells: AKT1(Tyr474), AKT2 (Ser474), ERK1-p44/42 MAPK(Tyr204), mTOR(Thr2446), p38 MAPK(Thr180), Rac1/cdc42(Ser71), SAPK/JNK(Tyr185) and SP1(Thr739) (2).

Discovery of so many signaling molecules was only possible as the antibody arrays allowed the screen dozens (or even hundreds!) of targets in a simultaneous way. Probably, if the research had involved picking different targets and doing individual WBs, there would have been a byass on what markers to study, and some of the ones mentioned above may never have been detected.

Th17 cells also have a role in tumour microenvironment (TME).  It has been found that the Th17 cell survival factor, IL-23, is overexpressed in tumor tissues isolated from mice and human breast cancer patients. It has been indicated that tumor-secreted PGE2 induces IL-23 production in the TME, leading to Th17 cell expansion. This inductive effect of PGE2 is mediated through cAMP/PKA signaling transduction pathway (3).

For further updates on effector molecules in the Th17 pathway, don’t hesitate to suscribe to our blog!

References

1.- Korn, R. et al (2009). Annual Review of Immunology. Vol. 27: 485-517. DOI: 10.1146/annurev.immunol.021908.132710.

2.- Hasan, M. et al (2015). Immunol Cell Biol. 2015 Aug;93(7):662-72. doi: 10.1038/icb.2015.21.

3.- Qian, X. et al (2013). J Immunol. 2013 Jun 1;190(11):5894-902. doi: 10.4049/jimmunol.1203141.

Patient-derived tumour organoids (mini colon tumours). In blue: cellular nuclei; in red: cellular membranes (Image: Alexandre Calon, IRB Barcelona).

Researchers at the IRB in Barcelona have found a signature of 4-6 genes able to predict the aggressivity of colon tumours, by analysing the tissue surrounding the tumour cells.

The scientists are currently developing a test that enables the identification of patients at risk of relapse after surgical removal of the tumour by measuring these found genes. They also propose to test in patients a particular drug that blocks the metastatic capacity of colorectal cancers in mice. This drug has been already tested using organoids derived from patients’ samples.

Source:

• Discovery of the genetic fingerprint of aggressive colon tumours – IRB Scientific news.
• Calon A et al. “Stromal gene expression defines poor-prognosis subtypes in colorectal cancer” (2015) Nat Genet Apr;47(4):320-9. DOI: 10.1038/ng.3225.

On a more funny note, have a look at the video done this last summer by researchers at the IRB Barcelona!

Any exciting research (or videos!) done over at your laboratory or institute?

We would like to know!

Research performed by David García-Molleví and his team at ICO (Barcelona, Spain), in cooperation with tebu-bio laboratories, is being presented at the AACR congress in Philadelphia these days.

The poster describes how cytokine profiling of drug-disrupted tumour cell / fibroblast crosstalk provides insights to understand the protective role of the stroma. Briefly, an array including 174 cytokines was performed on tumour cell / fibroblast co-cultures. Results determine that IL1b and TGFb1 secreted by tumor cells trigger the activation of normal colonic fibroblasts (NCF) to become CAFs. The role of IL1b is not as well known as TGFb1 in a cancer context.

Cytokine arrays were used in order to determine:

i) cytokine profiling of IL1b-treated NCF
ii) profiling of tumor cell-NCF cocultures in the presence of inhibitors of IL1b and TGFb1 signaling, main triggers of NCFactivation.

This poster shows how cytokine profiling can be useful as a complementary approach for microenvironment studies in assessing reciprocal activation of tumour cells and stroma, mediators of such interplay, treatment effectiveness and new target interventions.

Would you like to have a copy of this poster? Contact us!