At AstraZeneca, we are dedicated to changing the future for CKD patients. While there are currently available treatments that benefit patients with CKD, there are no treatments that specifically target the cause of CKD. We are investigating the drivers of disease and progression of CKD using a rigorous, science-led approach and adopting techniques that aim to break new ground in research. Our ambitions are high: we want to stop disease progression, manage morbidity and mortality, and ultimately modify and even reverse the disease itself.
By focusing on early disease detection underpinned by precision medicine, we hope that in the future patients will be matched with treatments most likely to work for them.

The kidney has critical roles in the human physiology, removing waste products and balancing the body’s fluids. It is also involved in metabolism and vitamin D production, and produces hormones that regulate red blood cell production and blood pressure.⁴ This all happens as a result of complex interplay with other organs, so CKD has knock-on effects throughout the entire body.⁵

For these reasons, we take a holistic view of the patient and are guided by the commonalities between these diseases in our research.
 

This is an exciting time – recent advances in three important areas lay the foundations for a new era in CKD research:

• Bioinformatics and ‘omics analysis to classify patients and uncover new genetic disease drivers

• The development of advanced models for target validation and understanding disease development

• New modalities for previously undruggable targets

CKD encompasses various primary disorders and stages of progression, and the patient population is highly heterogenous – CKD is really an umbrella term for many diseases. The current symptom-based approach ignores the different underlying molecular causes of disease and leaves potential for misdiagnosis. We aim to close this gap: enrolling the right people for the right trial is critical for research success and patient health.  

In the age of ‘omics and data processing, we believe we have reached a turning point. We are working collaboratively to build – and make sense of – huge genetic datasets from real patient samples.

This approach means we are uncovering different underlying molecular disease profiles. We hope then to be able to classify patients more accurately, and identify new biomarkers and disease targets.

Inspiration can sometimes come from unexpected places. Anna Reznichenko, Principal Scientist, Research and Early Development, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, has devised a novel method based on market segmentation analysis for handling massive datasets drawn from the financial industry. This approach has revealed distinct new patient categories, and could be used as the base for precision medicine in the future. 

Using data from the Renal Pre-competitive Consortium (RPC2) – a unique partnership of industrial and academic collaborators – gives Anna access to the largest-ever patient sample bank for renal transcriptomics (analysis of gene expression in kidney biopsies), containing over 250 genome-wide expression profiled samples, with data from patients at all stages of disease. Distinct, homogeneous patient categories can be most effectively revealed using a dataset of this size. However, access to the data itself is only part of the picture: the next step is analysing the patterns within the data points.

Using the unique datasets, the team applied the above described machine learning and artificial intelligence algorithm to classify patients into homogenous subclasses. For the first time, distinct disease categories based on molecular data which are different from previous clinical classifications for CKD could be seen. We are now looking to identify urinary biomarkers which could be used to reveal patients’ molecular disease classes non-invasively, allowing us greater precision when allocating the right patients to the right trials and in the future, potentially provide tailored treatments based on scientifically-determined individual disease categories.  At AstraZeneca, we are focused on regenerating the kidney as well as improving the distribution of blood and the supply of oxygen to kidney, and we have stem cell approaches focused on these areas. With our expertise in production of miniature organs (organoids) for testing new drug modalities, we are also exploring the tantalising prospect of growing kidney organoids as a therapy option for CKD – an approach arousing considerable interest among our researchers committed to improving the outlook for patients with CKD.¹

In collaboration with Columbia University, led by Professor David Goldstein and Professor Ali Gharavi, we have conducted the largest-ever exome sequencing study of CKD patients. The research results have given us valuable clinical insights into the genetic causes and therapeutic opportunities for this condition.

Published in the New England Journal of Medicine, the study is based on the results of exome sequencing more than 3,300 patients. It identified potentially causative genetic variants for a significant number (~9%) of patients. The research also revealed that six genes collectively account for almost two thirds of genetic diagnoses, and genetic diagnoses could be assigned to ~17% of previously uncharacterised CKD cases. This research could represent a significant breakthrough for those patients with a genetic diagnosis, as these data could potentially provide novel clinical insights to help inform treatment and care options.

These data enhance our disease understanding, as well as suggesting that genetic diagnosis can now be used in patients with previously unknown causes of the disease, and highlights the potential of early genetic testing to accurately identify and target patient subpopulations towards relevant clinical trials and targeted therapies.

We are in a stronger position than ever before to look for targets and treatments now we have a deeper molecular understanding of CKD. To use this new knowledge to our best advantage, we need the right research tools to simulate the disease and test our hypotheses. We are investing in the development of new, sophisticated models that mimic the human kidney more accurately than has ever previously been possible.

The complexity of the kidney means it has been virtually impossible to emulate in classical in vitro systems, while in vivo models are resource intensive to produce, and not always translatable to human systems. 

The new model systems we are embedding into our research pipeline to bridge this gap include 3D bioprinting and organs-on-chips. With these groundbreaking techniques, we can test compound behaviour and hope to shed new light on previously elusive disease mechanisms.

3D bioprinting

A 3D model with the vascular and tubular aspects of the kidney to emulate cell cross-talk in different organ compartments is the goal that until now may have seemed beyond scientists’ reach. However, we have moved a step closer towards modelling the human system by using 3D bioprinting in our collaboration with world-leading experts at Harvard University.

3D bioprinting involves printing-like techniques, where fugitive ‘ink’ is printed onto a gelatin/fibrogen matrix to pattern open vascular and tubular channels. These are then populated with cells to create biological structures. Organ segments can be created that preserve the architecture and heterogeneity of their biological counterparts – major progress from 2D modelling.

We have already bioprinted a 3D model with both kidney tubules and blood vessels, which replicates the behaviour of the two structures and importantly the interactions between the different tissue types. Modelling this interplay, which would take place in a real, living kidney is crucial for the creation of a realistic, relevant model system.

Creating realistic, living components of a healthy kidney will be a vital tool for understanding kidney function. As our aim is also to understand kidney dysfunction, we also are embarking on an ambitious project to create kidney components with diseased cells, using human induced pluripotent stem (iPS) cells from real patients. Ultimately, the logical conclusion of this work would be to print all the components needed to create an entire synthetic kidney. Although we are not at this stage yet, it is not outside the realm of future possibility.

Organs-on-Chips Technology

Organs-on-Chips technology is a game changer for scientists at AstraZeneca: these micro-engineered systems model the in vivo micro-environment, and represent a new paradigm for cell-based models. We are using them to model the glomerulus – the kidney’s filtration barrier – which breaks down in CKD. Why this happens is still a mystery; we hope this technology might help bring some answers.

Until now, 2D cell-based models used for compound testing have not been able to reflect the complexity of the kidney environment and results would not always be a faithful representation of what happens in the body.

Observing a complete 3D system enables a better understanding of kidney function – and dysfunction.  Kidneys are made up of over 20 different cell types, arranged within an intricate architecture. Crosstalk between these cells and their interaction with their micro-environment drastically influences their behaviours in normal function, disease processes, and how they respond to potential drugs.

Organs-on-Chip technology provides an environment that emulates key aspects of the physiology of a real kidney; including fluidic channels that are lined with living cells in the correct architecture. The flow represents the flow of blood carrying nutrients, takes away waste products, and also creates mechanical forces, all critical factors for the functioning of the kidney.

3D kidney microphysiological systems (kidney-on-chips) are enabling different cell types to be co-cultured, allowing cellular interactions and signalling to be investigated in both healthy and diseased kidneys.

With our partner, Emulate, Inc., we are developing Organ-on-Chip models to demonstrate the utility of this technology as a more predictive alternative for efficacy and safety testing of new chemical entities. For the first time, the growth and interaction of three different cell types which reflect the filtration barrier of the kidney have been studied using glomerulus-on-a-chip technology. Initial findings showed that the three cell types can successfully cross-talk with each other, creating a highly physiologically relevant tool to study healthy glomerular function. We hope it will enable greater understanding of the mechanisms underlying chronic kidney disease and improved assessment of new therapies. Read more about our research into Organs-on-Chips here. 

Antisense oligonucleotides

The next step on from our breakthrough genetic research is identifying drugs that target the root causes of kidney disease, recruiting patients with specific genotypes into relevant trials where they are likely to gain greatest benefit.

In partnership with Ionis Pharmaceuticals, we are researching APOL-1 – a gene with variants that is related to an increased risk of early onset kidney disease and rapid progression. People of West-African ancestry are at increased risk of developing end stage renal disease, and common APOL-1 polymorphisms may account for this risk.⁶

Understanding that APOL-1 is a genetic driver of disease leads us a step closer to addressing a high unmet need in a defined population. If we can target the high-risk APOL-1 variants, this could lead to potential treatments for people who have limited options.

One emerging treatment strategy is to target protein expression using antisense oligonucleotides (ASOs). These are short chains of nucleotides that bind to mRNA, thereby modulating protein expression.  ASOs can be designed to bind to almost any defined genetic sequences, meaning they have potential for use in diseases with genetic origin.

Cell therapy

Expanding knowledge on the cells involved in CKD is allowing greater precision in identifying and reaching relevant therapeutic targets. Our latest research is exploring the potential of renal progenitor cells (a type of stem cell) to replace lost or damaged renal cells. When delivered either intravenously or within the kidney itself, these progenitor cells can differentiate and replace cells lost to disease, an exciting prospect for next generation therapeutics.

In addition, we are applying our cellular models that accurately mimic the function of the kidney as potential new therapeutic options for CKD.  Organoids which comprise at least 11 different cell types that are needed for the full organ to function normally, have already shown high fidelity to the whole human kidney. These translatable systems are enhancing our understanding of kidney dysfunction and providing new approaches and technologies for treating CKD.

Find out more about our approach to cell therapy in damaged kidneys.