Visit to Immundnz in February 2018

I recently had the pleasure of visiting Immundnz, a young company based at Alderley Park in Cheshire. My hosts were the two founders of the company, Dr Masih Alam and Dr Robert-Jan Lamers.

The company was founded in 2016 to provide specialised support in immunology. They use an array of cellular, immunologic and molecular methods to build up a comprehensive picture of drug-induced immunogenicity.

Masih is a PhD immunologist from the University of Manchester and was able to explain current immunological thinking to me in terms understandable to an analytical chemist - although it did require more than one cup of coffee!

Robert-Jan is an Analytical Chemist with a PhD in Bioanalytical Chemistry from the University of Leiden and great experience in separation science and mass spectrometry.

The Theory

Masih explained how the contemporary theory of the immune response recognises the importance of homeostasis and that that the immune system is not programmed to respond to foreign particles, as it was conceived for many years during the “self-non-self” era of immunology. What turns the immune system on and off is a very important question in understanding disease and pathology. Furthermore, it is the condition of the tissue that is central to understanding homeostasis, tolerance and the immune response. The central dogma of immunology now is that the immune system receives signals to respond from the ‘tissue’ and that the immune system is far less concerned with things that are foreign than with those that do damage.

So, tissue damage is very central to inflammation and immunogenicity and understanding tissue damage is pivotal to assessing immunogenicity and pathological risk. In fact, any drug molecule has the capacity to cause unwanted tissue damage and the outcome of such damage would be the release of endogenous molecules to the immune system. An unwanted immune response may be generated from such signals and that may progress into a secondary disease including autoimmunity.

Consequently, it is upon this tissue damage, as the central driver of an immune response, that Immundnz focuses. Whether the damage is an outcome of a drug molecule, a pathogen or endogenous metabolites, the prediction of a drug molecule’s immunogenicity or cytotoxicity to non-target tissue will be best understood at this upstream event...

Some well-known examples

Masih explained how severe tissue damage caused by the marketed antibiotic Trovafloxacin led to its withdrawal from the market with great financial and human costs. The lethality of the drug was found to be through causing necrosis of hepatocytes leading to liver failure. The importance of anything that can help avoid repetitions of such tragic situations is blatantly obvious.

Most people will also have heard of the story of TGN1412, an immunotherapy developed (briefly) by a TeGenero in 2006 which led to the infamous Phase I incident in the UK. But there are many drugs on the market that are known to cause secondary disease. In many cases it is not known why the secondary adverse effect happens. The use of these drugs, in some cases on a wide scale, is due to their greater benefit of curing a life-threatening condition. Autoimmunity is a common secondary pathology, and drug induced lupus (DIL) accounts for 10% of all lupus incidences. Here are a few more examples of drug side effects:

• Penicillin – autoimmune haemolytic anaemia
• Amiodarone – interstitial lung disease
• Adalimumab – DIL, vasculitis
• Erythropoietin – red blood cell aplasia, ADA
• Pegferon- hyperthyroidism
• Carbamezepine – DIL

Conventional ‘immunogenicity’ testing considerations are limited to ‘antigenicity’ testing of the drug molecule only. In standard toxicology studies, immune response data such as infiltrations of lymphocytes in tissue sites are ignored as transient immunity and the possibility of disease occurring from chronic immunogenic events is overlooked. And it is this broader immunogenicity of a drug that Immundnz considers and their assays are designed to answer questions about drug-induced tissue damage and immune response.

Facilities and Services

Masih and Robert-Jan gave me a comprehensive tour of their laboratory facilities so I was able to see at first-hand how they support their service portfolio with various techniques and methods. In immunology, they have a range of methods and techniques available such as microscopy, flow cytometry, fluorescence and absorbance reading, western blotting, immunohistochemistry and  histology.

 One of Immundnz's labs

One of Immundnz's labs

Cell culture systems are at the heart of their expertise and services. They grow human and mammalian cells in various culture media, both adherent and non-adherent (e.g. lymphocytes) type. They grow established (cell lines) and primary cells in 2D cultures as well as in 3D tissue models such as organoids and 3D microfluidic culture plates. These in vitro systems are used to investigate biological features and drug compound interactions. Some frequently used cell types in the laboratory are HEK293, HeLa, HepG2, Thp-1 (monocytic), human aortic SMC, fibroblasts, dendritic cells, T cells and B cells.

Crucially, high-end mass spectrometry is provided by sister company Abundnz based in The Netherlands where they specialise in the characterisation of cells, cell fluids and culture supernatants. The endogenous molecules or proteins that are released by cells through drug-induced necrosis or compromised membranes are activators of an immune response.  By characterising these molecules in cells and cell fluids in combination with pathway analysis and sophisticated bio-informatics, they are able to assess drug-induced tissue damage.

COMPIT

So, as we’ve seen, immunogenicity of drugs is a very serious issue and can lead to massive complications with financial loss. Immundnz brings new insight to this term from an immunology point of view. COMPIT, Comprehensive In Vitro Human Immune Response Testing of drug compounds, is their main product that studies the comprehensive immunogenicity of a drug. They are currently conducting an international grant project to research and develop a platform that enables the preclinical study of drug induced immunogenicity and the risk of immune mediated pathology. It has been designed to enable preclinical in vitro testing of drugs for an in-depth immunogenicity analysis – currently not offered by other established analytical systems. The system is based on the central dogma of immunology and the immune response against ‘Tissue Damage’.

COMPIT merges cutting-edge immune response testing via a system of biological assays combining novel cell lines, 3D tissue models and primary cells to deliver an in vitro human cell-based immune system that recapitulates the in vivo human microenvironment and can be characterised by ultrasensitive analytical assays.

COMPIT outperforms current immune response testing and is complementary to current information obtained from preclinical studies. It can be used during preclinical development to assess drug-induced tissue damage and immunogenicity and to predict the human immune response as a result of a drug and drug-induced tissue damage. As such, COMPIT is designed to fit into the current scheme of preclinical studies and can be applied to both small and large molecules. Particularly the research and development of large molecules is evolving and expanding worldwide and requires a better and more in-depth insight into the human immune response.

COMPIT brings numerous benefits:

• Prediction of immune-mediated adverse drug reaction and avoiding risk in the clinical phase
• Better translation from preclinical studies to humans
• Reduction of the number of animal tests
• A more robust and comprehensive immunologic profile: improved safety and risk prediction thus reducing human health hazard risk
• Drug development can be terminated before costly advanced stages of drug development: reduced investments in non-viable drugs and reduced attrition rate
• Potential to improve drugs, generate blocking agents against immune effectors and make non-viable drugs accessible to patients
• Meeting changes to expected regulatory requirements on safety

Further Information

If any of the issues discussed in this article are of interest to you, please feel free to contact me directly on mark@discoveryanalyticalconsulting.com for further information.