What do NGS and CRISPR-Cas9 mean for the future of drug products?
By: Kristie Somers, BSc.
Researchers developing new therapies are practicing novel scientific techniques to better identify, prevent, and treat diseases. The transition from blanket therapeutic approaches to customized treatments for a single patient is referred to as precision medicine. Many pharmaceutical companies are developing treatments based on the precision medicine model as it reduces trial-and-error prescribing and emphasizes prevention rather than reaction.
At Vanrx we work to continue our innovation and determine new applications for our Aseptic Filling Workcells. Therefore, as new scientific treatments are developed Vanrx is able to design products for these next generation therapies. Biologics came about as a result of genomic sequencing, which now leads to cell & gene therapies and gene editing. The result of this is a shift towards therapies becoming more personalized. As we understand this shift, we are able to help bring new drug products to market quickly, which will provide a positive influence on human health.
Before I joined the Vanrx team, I was in a technical sales role which allowed me to visit dozens of labs engaging in medical research. From this exposure and my biomedical background, I gained experience with many gene editing therapies and techniques. The objective of this article is to discuss two scientific tools and how they influence the future of precision medicine: Next-generation sequencing (NGS) and CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats).
Next-Generation Sequencing (NGS): determining the genetic profile
Precision medicine uses patient genomic data to help provide the right treatment for the right patient at the right time. NGS technology is a diagnostic tool that allows for the rapid and accurate sequencing of many genes of an individual patient for early detection of disorders. NGS is frequently used in forensics and for routine detection of many genetic diseases including; infectious diseases, immune disorders, human hereditary disorders, and therapeutic decision making for somatic cancers.
Each country has their own set of guidelines governing the NGS process however, they all categorize NGS as a diagnostic test – a test directed toward answering a clinical question related to the genetic structure and potential medical condition of a patient. The process is similar to that of a blood test, however the sample collected from the patient can be any bodily fluid or tissue. The sample is then delivered to an NGS core facility or clinical lab that specializes in NGS sequencing. Commonly NGS testing only targets the exome — the 2-3% of the human genome that codes for proteins and yields ~85% of disease trait mutations. This is referred to as whole-exome sequencing (WES) and is currently used as a standard diagnostic approach for the identification of molecular defects for suspected genetic disorders, such as Miller syndrome.
The initial step of an NGS workflow – library preparation – is the most critical. The library preparation, which depends on the trait being investigated, alters the patient’s RNA or DNA fragments into a form that is readable by the sequencer, ie. NextSeq or MiSeq platform. Once the material is sequenced, the sequencer will provide data that reflects the genetic make-up of the patient. This genome is compared to that of a standard using comparative genomics software. A rating system is used to report on all NGS-based diagnostic tests performed. The report will summarize the patient’s identification and diagnosis, a brief description of the test, a summary of results, and the major findings. This information is relayed to the patient and if there is a genetic disorder seen in the results, further steps for treatment are taken. In terms of precision medicine, these treatments include cell & gene therapy, CAR-T, oligo synthesis, and CRISPR-Cas9.
What does NGS mean for the future of drug products? NGS sequencing technology has transformed genomic research to the point were many researchers that I have had the privilege of learning from are on the brink of discovering the genetic codes and subsequent therapies for many different cancers and neurological disorders. In the future, NGS might be a common test performed when you are born. This way your genetic profile can be recorded and any changes in your genetic code via mutations can be tracked. The earlier we can detect diseases, the quicker we can provide a therapy.
CRISPR-Cas9: gene-editing cell therapy
CRISPR-Cas9 gene-editing is a versatile therapeutic platform for genome editing to treat diseases such as cancer, cystic fibrosis, muscular dystrophy and sickle-cell anemia. CRISPR-Cas9 allows scientists to make precise changes to the genetic code of living organisms. The acronym stands for ‘clustered regularly interspaced short palindromic repeats’. Cas9 is the ‘CRISPR-associated protein molecule’ that performs the cutting action. Literally speaking, CRISPR-Cas9 is a ‘cut and paste’ tool for editing gene sequences.
Once a condition is diagnosed through NGS or other means, the CRISPR-Cas9 process begins with extraction of cells from the patient which are transported to a clinical laboratory. Using CRISPR-Cas9, the cells are modified to remove the gene that encodes for a receptor with the mutation. This is done using a specially designed guide-RNA molecule that leads Cas9 endonuclease to the target DNA, which Cas9 will cut. In response to the cut, an endogenous repair mechanism is triggered, which will either generate a gene knockout (remove the gene) or introduce a modification of DNA with the desired sequence. For example in cancer treatments, T cells would be extracted from a patient, and genetically modified using CRISPR-Cas9. When the ‘new’ T cells are reintroduced to the patient via continuous IV injection, the are ‘super charged’ meaning they have a higher capacity to attack tumor cells.
What does CRISPR-Cas9 mean for the future of drug products? Although gene-editing has varying levels of acceptance and is still in a development stage, CRISPR-Cas9 is a key breakthrough for precision medicine. In China there are several successful clinical trials for many different forms of cancer being treated with CRISPR-Cas9. Also, clinical trials are underway in the USA for CRISPR-Cas9 as a treatment for sickle cell disease and is being explored for the treatment of cystic fibrosis. These breakthroughs mark the first step in what could become a common way to treat genetic diseases. The full potential of this gene-editing tool is being explored further and is a stepping stone to the arrival of precision medicine.
NGS and CRISPR-Cas9 have heavily influenced the progression of precision medicine. The ability to understand a patient’s genetic profile has provided enough information to use gene therapies and genetic manipulation for personalized treatment. Vanrx is a company that is aware of the scientific advancements in medicine and supports growth. By remaining deeply connected in the industry, the company continues to develop new tools to meet the changing needs of pharmaceutical companies. We are continuing to improve existing systems and designing for the therapies coming next.