White Paper – Use of Next Generation Protein Therapeutics Over Traditional Monoclonal Antibodies and Different Approaches


USE OF NEXT GENERATION PROTEIN THERAPEUTICS OVER TRADITIONAL MONOCLONAL ANTIBODIES AND DIFFERENT APPROACHES

A recent white paper I developed for Informa.

Available now on knect365.com. Click the link to access the white paper.


Increasingly next generation proteins are being used over traditional monoclonal antibodies. This exclusive whitepaper explores the different approaches, competitive advantages and challenges of these next gen therapeutics.

According to BIS Research and Research and Markets the worldwide biologics drug discovery market is predicted to increase from USD $8.1 billion in 2015 to USD $22.7 billion by 2025. The biologics discovery growth rate is predicted to be faster than in the dominant small molecule discovery sector. Monoclonal antibodies are expected to make up just under half of the biologics discovery market in 2025. Engineered or recombinant proteins such as next generation protein therapeutics are expected to contribute 3 billion to the market in 2025.

The dominance of monoclonal antibodies (mAbs) are due to several factors including their efficacy brought about by their high specificity allowing selective targeting; long half-life compared to small molecules due to the presence of the Fc region that allows the antibodies to engage the FcRn mediated salvage; and the ability to activate immunemediated effector functions, e.g. antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), via the interaction of Fc portion with Fc-gamma receptors. Promoting their adoption, Mabs have proven useful in the rapidly expanding immune-oncology field which has led to a proliferation of antibody products through “copy-cat” development. Mabs, however have important disadvantages such as cost of goods that next generation protein therapeutics do not suffer from.

Next generation protein therapeutics can be considered antibody mimetics as each protein therapeutic is composed of a constant region, which stabilizes the overall protein folding, and variable regions that facilitate its target binding. The specificities of these next generation proteins variable regions can be directed in vitro, contrasting antibodies and antibody fragments which require the immunisation of animals to generate.

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Next Generation Treatments For Type I Diabetes

Note this article was originally published on apptheneum on July 15, 2014. 

 

Type I diabetes also known as juvenile diabetes is an autoimmune disease primarily mediated by T-cells (a type of white blood cell) that results in the destruction of pancreatic β-cells which produce the body’s insulin hormone. Insulin induces the uptake of glucose by adipose tissue (fat cells) and skeletal muscle. Lack of insulin results in hyperglycaemia (higher than expected blood glucose). The three major symptoms of hyperglycaemia are frequent acute, hunger and thirst, and increased volume of urination. If untreated coma and heart attack can result. Standard treatment is daily insulin injection. This does not perfectly regulate blood glucose levels and over time people with type I diabetes can develop problems with their feet associated with reduced circulation and nerve damage. Cardiovascular disease, retinopathy (eye damage), general nerve damage, kidney disease, and sexual dysfunction are also major problems. The next generation of gene and cell therapies aim to restore glucose sensitive insulin secretion in type I diabetics and thus eliminate long-term complications.

 
Cell therapy offers much promise but has yet to prove it can overcome certain obstacles. It is based on replacing the destroyed β-cells with either mature donor cells or (theoretically) autologous cells generated from the patient. Unfortunately the underlying disorder reamains and the cells will eventually be destroyed by the immune system once more even with the administration of immunosuppressive agents. This approach also suffers from lack of donors. None the less the initial transplantation method has been proven effective. Human Embryonic stem cells (from donors not the patient) have been used to generate insulin secreting β-like cells. However immunosupression is still required to administer these cells and they suffer from the same long term degradation as donor islets.

 
Patient derived cells are potentially a better option as they do not require a donor and would not trigger an anti-self immune response when reintroduced to the patient. Type I diabetes patient derived induced pluripotent stem cells (iPS) have been generated from skin biopsy fibroblasts and insulin-producing and glucose-responsive β-like cells have been differentiated from these cells. Minicircle DNA vectors for eukaryotic cell transfection lack any bacterial or viral sequence. In contrast to standard vectors that contain bacterial sequences transgene expression from minicircle vectors can be sustained for weeks. They also have the advantage that they do not integrate into the host cell genome which eliminates concerns of tumourigenic insertion events. Minicircle vectors have been used to generate iPS cells. This provides a basis for the development of treatments for type I diabetic patients based on transplantation of β-like cells differentiated from these minicircle generated iPS.

Fig1-glucose uptake

 

Gene therapy offers a potentially less complicated method of restoring glucose regulated insulin secretion to type I diabetic patients. Furthermore this approach has proven effective long term (reversal of diabetes for greater than 4 years) in a large animal model (induced diabetic Beagle dogs). Callejas et al. targeted skeletal muscle for insulin production. Insulin stimulates both the translocation of GLUT4 to the plasma membrane and the activity of hexokinase (Figure 1). Glucokinase is analogous to hexokinase but is more constitutively active but requires high glucose concentration for maximal activity. In the Callejas et al. study both insulin and glucokinase driven by constitutive promoters was delivered to the dog’s muscle by injection of adeno-associated viral vectors. In these animal hyperglycemia was eliminated and remained so for greater than four years (until the end of the study). There was no evidence of diabetic complications in these animals. The authors aim to replicate these results in pet dogs and eventually human clinical trials. If successful type I diabetes could be treated with a one time procedure that would afford long term protection from diabetic complications.

 
The liver is an attractive target for type I diabetes gene therapy due to its relative ease of targeting with non-viral vectors in animal models (hydrodynamic injection) and the fact that hepatocytes (liver cells) have an active protein synthesis and constitutive protein secretory mechanism, as well as the ability to sense and metabolically respond to changes in ambient glucose levels. In a recent study minicircle DNA containing modified insulin gene to facilitate liver processing under the control of glucose response elements was administered to chemically induced diabetic rats via hydrodynamic injection. Hyperglycaemia was eliminated for a month following the initial hydrodynamic procedure. This approach is attractive as it is potentially very safe with no DNA integration and no bacterial or viral sequences to trigger immune response. However a delivery method based on the mechanism of hydrodynamic injection needs to be developed for application to patients in the clinic.

 
The endocrine K cells of the gut are an attractive target as they are glucose responsive and secrete the hormone glucose-dependent insulinotropic polypeptide (GIP) in response to glucose in a temporal pattern very similar to insulin. It has been demonstrated that when the GIP promoter is used to drive insulin expression and introduced into the pronuclei of fertilized mouse embryos, the transgenic mice that are generated express insulin from their K cells. Furthermore when the transgenic mice were treated with Streptozotocin a β-cell toxin they did not develop diabetes in contrast to control mice. On the basis of this research the Canadian company enGene are currently developing methods to target human K cells for type I diabetes therapy.

 
Gastric G cells, exocrine pancreas and L cells have also been targeted. Thus, there are many options awaiting clinical trial. However a major obstacle to all of the approaches mentioned which is often not accounted for in the studies discussed above is the original autoimmune disease of the patient that caused the destruction of the β-cells in the first place. Fortunately progress has been made in understanding the underlying immune response that triggers type I diabetes however much work is needed to determine an approach that would suppress the autoimmune response in diabetic patients. A very interesting observation and one that could be used to protect novel reprogrammed β-cells is that the adenovirus E3 gene cassette can protect β-cells from destruction in a virus induced mouse model of diabetes.

 

Further clinical trials for type I diabetes are required to determine which of these promising experimental treatments is the best option for this autoimmune disease.

 

David Orchard-Webb

 

Bibliography

Alam, T. et al. Correction of diabetic hyperglycemia and amelioration of metabolic anomalies by minicircle DNA mediated glucose-dependent hepatic insulin production. PloS one 8, e67515 (2013).

Auricchio, A. et al. Constitutive and regulated expression of processed insulin following in vivo hepatic gene transfer. Gene therapy 9, 963-971 (2002).

Callejas, D. et al. Treatment of diabetes and long-term survival following insulin and glucokinase gene therapy. Diabetes 62, 1718-1729 (2013).

Chen, Z.-Y., He, C.-Y., Ehrhardt, A. & Kay, M.A. Minicircle DNA vectors devoid of bacterial DNA result in persistent and high-level transgene expression in vivo. Molecular therapy 8, 495-500 (2003).

Cheung, A.T. et al. Glucose-dependent insulin release from genetically engineered K cells. Science 290, 1959-1962 (2000).

Jia, F. et al. A nonviral minicircle vector for deriving human iPS cells. Nature methods 7, 197 (2010).

Maehr, R. et al. Generation of pluripotent stem cells from patients with type 1 diabetes. Proceedings of the National Academy of Sciences 106, 15768-15773 (2009).

E. Tudurí et al. Reprogramming gut and pancreas endocrinecells to treat diabetes. Diabetes, Obesity and Metabolism 13, 53-59 (2011).

Von Herrath, M.G., Efrat, S., Oldstone, M.B. & Horwitz, M.S. Expression of adenoviral E3 transgenes in β-cells prevents autoimmune diabetes. Proceedings of the National Academy of Sciences 94, 9808-9813 (1997).

 

KRAS, CDKN2A, SMAD4, and TP53 Gene Mutations Linked to Pancreatic Cancer Patient Survival Time

Alterations in four main genes are responsible for how long patients survive with pancreatic cancer, according to a new study.

via Pancreatic cancer survival linked to four genes — Pancreatic Cancer News — ScienceDaily

The Medical Laboratory Scientist Personnel Shortage And The Future Of Lab Medicine

 

It was recently reported that the United States molecular diagnostics market is expected to exceed $9 billion by 2024 due to the increased incidence of infectious diseases and different types of cancer. In fact, there are at least 4 billion laboratory tests performed in the U.S. each year used to generate data on these diseases.

 

As this market continues to increase, so does the need for medical laboratory scientists. Correspondingly, there is a 22 percent projected growth of medical laboratory jobs that are needed to help physicians and patients detect and treat diseases faster. The United States’ population aged 65 and over will be comprised of around 83.7 million citizens in 2050. This increase in the aging population will lead to a greater need to diagnose medical conditions, such as cancer or type-2 diabetes, through laboratory procedures.

 

To learn more about the growing molecular diagnostics market and the future of medical laboratory science, check out the graphic below created by the University of Cincinnati.

 

MedicalLab-Scientist-Shortage-NEW

 

New Publication: Biobanking Weekly

 

Click here to visit Biobanking Weekly

The inaugural issue of Biobanking Weekly is coming to you just as the Global Biobank Week is closing providing leaders at the cutting edge of Biobanking to set the agenda for the months ahead. Important developments in biobanking this week include contributions to understanding of type 1 diabetes, the risks of 9/11 responders to fall victim to human papilloma virus (HPV) related head and neck cancers, new targets for osteoporosis treatment, advances in banking prostate cancer samples, the use of cystic fibrosis biobanks, the testing of drones for sample delivery, and the adoption of lean sigma six for efficient management and distribution of samples. Very exciting developments with plenty more forecast for the coming weeks.

Curcumin analogues extracted from Alpinia officinarum and Alnus japonica inhibited the FoxM1 signalling axis in a pancreatic cancer cell line

 

Alpinia officinarum or lesser galangal (高良姜), is a member of the ginger family, which originates from China and is now cultivated throughout Southeast Asia (Figure 1). The roots are known as galangal and are used in cooking, perfumes and are also known for their medicinal properties. Alnus japonica or East Asian alder (日本桤木), is a species of tree found in Japan, Korea, and eastern China, stretching to Russia. Diarylheptanoids, the family of which the anti-cancer agent curcumin from turmeric is a member, can be extracted from these plants. Diarylheptanoid compounds from these medicinal plants were found to inhibit the growth of the PANC-1 (KRAS heterozygous G12D, TP53 homozygous P72R and R273H) pancreatic cancer cell line [1, 2]. The mechanism was proposed to derive from inhibiting the FoxM1 transcription factor signalling axis.

 

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Figure 1: Alpinia officinarum. Credit: Biodiversity Heritage Library. No changes were made. Creative Commons Attribution 2.0 Generic (CC BY 2.0).

 

FoxM1 is a transcription factor of central importance to pancreatic cancer [3]. It promotes the transcription of genes involved in cell cycle progression and cell survival as well as migration and invasion [4]. FoxM1 transcription has been demonstrated to be promoted by the sonic hedgehog pathway in colorectal cancer and furthermore the hedgehog pathway is almost universally upregulated in pancreatic cancer [5, 6]. Dong et al. proposed that FoxM1 target genes are downregulated in response to the diarylheptanoid compounds due to Gli1/2 protein downregulation. Interestingly Stat3 signalling has been found to be downregulated by other diarylheptanoid compounds such as HO-3867 and Stat3 transcription can be indirectly upregulated by Gli1 via IL-6 [7, 8]. The extent to which different diarylheptanoid compounds could inhibit both FoxM1 and Stat3 axes in pancreatic cancer  is an open question.  

 

Refs

  1. Dong GZ, Jeong JH, Lee YI, Lee SY, Zhao HY, Jeon R, Lee HJ, Ryu JH. Diarylheptanoids suppress proliferation of pancreatic cancer PANC-1 cells through modulating shh-Gli-FoxM1 pathway. Arch Pharm Res. 2017 Apr;40(4):509-517. Doi: 10.1007/s12272-017-0905-2. PubMed PMID: 28258481.
  2. Gradiz R, Silva HC, Carvalho L, Botelho MF, Mota-Pinto A. MIA PaCa-2 and PANC-1 – pancreas ductal adenocarcinoma cell lines with neuroendocrine differentiation and somatostatin receptors. Sci Rep. 2016 Feb 17;6:21648. Doi: 10.1038/srep21648. PubMed PMID: 26884312.
  3. Akbari B, Mohammadnia A, Yaqubi M, Wee P, Mahdiuni H. Comprehensive Dissection of Transcriptome Data and Regulatory Factors in Pancreatic Cancer Cells. J Cell Biochem. 2017 Apr 12. doi: 10.1002/jcb.26053. PubMed PMID: 28401644.
  4. Quan M, Wang P, Cui J, Gao Y, Xie K. The roles of FOXM1 in pancreatic stem cells and carcinogenesis. Mol Cancer. 2013 Dec 10;12:159. Doi: 10.1186/1476-4598-12-159. Review. PubMed PMID: 24325450.
  5. Wang D, Hu G, Du Y, Zhang C, Lu Q, Lv N, Luo S. Aberrant activation of hedgehog signaling promotes cell proliferation via the transcriptional activation of forkhead Box M1 in colorectal cancer cells. J Exp Clin Cancer Res. 2017 Feb 2;36(1):23. doi: 10.1186/s13046-017-0491-7. PubMed PMID: 28148279.
  6. Jones S, Zhang X, Parsons DW, Lin JC, Leary RJ, Angenendt P, Mankoo P, Carter H, Kamiyama H, Jimeno A, Hong SM, Fu B, Lin MT, Calhoun ES, Kamiyama M, Walter K, Nikolskaya T, Nikolsky Y, Hartigan J, Smith DR, Hidalgo M, Leach SD, Klein AP, Jaffee EM, Goggins M, Maitra A, Iacobuzio-Donahue C, Eshleman JR, Kern SE, Hruban RH, Karchin R, Papadopoulos N, Parmigiani G, Vogelstein B, Velculescu VE, Kinzler KW. Core signaling pathways in human pancreatic cancers revealed by global genomic analyses. Science. 2008 Sep 26;321(5897):1801-6. Doi: 10.1126/science.1164368. PubMed PMID: 18772397.
  7. Hu Y, Zhao C, Zheng H, Lu K, Shi D, Liu Z, Dai X, Zhang Y, Zhang X, Hu W, Liang G. A novel STAT3 inhibitor HO-3867 induces cell apoptosis by reactive oxygen species-dependent endoplasmic reticulum stress in human pancreatic cancer cells. Anticancer Drugs. 2017 Apr;28(4):392-400. Doi: 10.1097/CAD.0000000000000470. PubMed PMID: 28067673.
  8. Mills LD, Zhang Y, Marler RJ, Herreros-Villanueva M, Zhang L, Almada LL, Couch F, Wetmore C, Pasca di Magliano M, Fernandez-Zapico ME. Loss of the transcription factor GLI1 identifies a signaling network in the tumor microenvironment mediating KRAS oncogene-induced transformation. J Biol Chem. 2013 Apr 26;288(17):11786-94. doi: 10.1074/jbc.M112.438846. PubMed PMID:23482563.