Pioneering a Neuvae™ Forward in Healthcare

25 min readMay 1, 2021

Solving Antimicrobial Resistance by Initiating an Immune Response Against Pathogenic Bacteria

There’s an old saying about death by a thousand papercuts. But what if it just took one papercut to kill someone?

That’s silly, right? I mean, even if a papercut DID get infected, we’ve got antibiotics to clean us right up…DON’T WE?

Behind the scenes, antibiotics have enabled much of modern medicine. We use them to cure infectious diseases, but also to safely facilitate everything from surgery to chemotherapy to organ transplants. Without antibiotics, even routine medical procedures can lead to life-threatening infections.

However, what if I told you that we’re living in a world where a papercut infection can be more deadly than cancer?

As time continues, bacteria continue to adapt and mutate to circumvent, identify, and destroy the threat of Antimicrobials. This is known as Antimicrobial Resistance, or AMR for short. When we use antibiotics, some bacteria die, but others survive and even multiply. The excessive and continued use of antibiotics allows for these “evolved” bacteria to become further resistant to the antibiotic. Thus making them extremely difficult or even impossible to treat.

Some bacteria can be so resistant that there are no antibiotics doctors can use to treat the bacteria. The term we use to define these resistant bacteria is known as “superbugs.”

Measuring the Impact.

AMR has emerged as one of the principal public health problems of the 21st century. Every year, 700k deaths occur solely due to Drug-Resistant Infections. As time continues, we can expect this to grow significantly. According to the CDC, this number will reach over 10 million deaths by 2050, with a 3.5% loss in GDP as a result of this crisis.

More than 2.8 million people in the US alone are already getting infected with AMR strain bacteria each year, and the cost to cure each patient with these strains has risen to 27k dollars.

Over the period 2015–2050, AMR will have cost the health systems of EU/EEA countries a total of USD PPP 60 billion, while in the United States, Canada and Australia, this amount will reach a combined total of approximately USD PPP 74 billion.

While developed countries have higher-end procedures and medical equipment, those in developing nations are 4x more likely to not just get infected, but also die of an AMR strain of a bacterial infection.

Each year, nearly two million people in the U.S. become infected with bacteria resistant to at least one antibiotic, and 23,000 of those people die.

And we can go on and on….

The Statistics Are Not Lying.

AMR is in Desperate Need of a Solution.

The Problem With Antimicrobial Resistance.

There is a race between the spread of AMR, and the development of new medicines, and it is a race that so far we are losing: AMR is growing, but new treatments are not. When formulating a solution to a problem of this magnitude, it is important to note the four main reasons why it exists.

The problem with Bacteria is that it is very easily able to swap genes the way we swap baseball cards thanks to a process called gene transfer, either sweeping up antibiotic resistance in the genetic remains of dead bacteria or exchanging it during a sort of bacterial makeout session we call conjugation.
Some bacteria like Staphylococcus have gained the ability to rebuild their cell wall faster than one antibiotic break it down. Other bacteria have “learned” how to make pumps that flush antibiotics out of the cell before they do their job.
Antibiotics we DO have today have come mainly from the environment, we’ve adopted the natural weapons that microbes use to wage war on each other… but that also means they’ve had billions of years to develop resistance. It seems like wherever nature has developed an antibiotic, it’s also developed a way to fight it. Resistance seems like an inevitable result of evolution. Of course, we are doing our part to help the superbugs succeed.
In some countries, people consume too many antimicrobials, often when they will not work. For example, it has been estimated that in long-term care facilities and general practices in the OECD area, up to 70% and 90% respectively of antibiotics are prescribed for inappropriate reasons. On the other hand, in other countries, people cannot afford to buy the drugs they need. In addition, antimicrobials are also heavily used in agriculture, often for no other reason than to make animals grow more quickly. Such ineffective use encourages AMR.

Now is the time to scale up global efforts to tackle AMR before it becomes uncontrollable

Introducing…Neuvae!

At Neuvae, our proprietary model harnesses the power of bacteriophages and transduction particles to produce an immune response against pathogenic bacteria and ultimately, eliminate the need for antibiotics.

We have spent countless hours creating something radical, something new, something that surpasses antibiotics ineffectiveness, solves the AMR crisis and most importantly prevents us from living in a world where a small cut could be more deadly than cancer.

Antibiotics are the fossil fuels of medicine. They are our only method of defence against bacterial infections in use right now, and their convenience in not just killing a lot of bacteria but also cost per bottle makes it ridiculously hard to find an alternative.

Our goal at Neuvae is not to provide an “alternative” to antibiotics. Instead, we want to create a 10x jump in the way that we treat bacterial infections so that our treatment methodology can be feasible and effective for the next decades to come and completely blow antibiotics out of the water.

Our Solution.

Neuvae’s solution consists of 3 pillars, each making 10x improvements in the traditional phage therapy pipeline.

The First Pillar — Proprietary bacteriophage engineering process that can reduce the cost of engineering by 80% while allowing us to engineer custom phages.
The Second Pillar — Administering the custom bacteriophage into the body and through its natural systems, target, disable and kill the resistant bacteria.
The Final Pillar — Boost the innate immune response to the same bacterial infection by using immunomodulatory particles carrying antigens transported through the phage.

Why Bacteriophages?

A bacteriophage is fundamentally a virus that exclusively targets bacteria. Whenever these phages target and lyse a bacteria, the clear area along with the remains of the lysed cells is known as a phage plaque. A simple way to understand is to imagine a tissue as a dense forest, with the cells being trees. A clearing in the forest, where there are no trees only some leaves and sticks on the group would be equivalent to the plaque. Due to the fundamental nature of bacteriophage lysis, the reproduced phages will always lyse outwards, developing several plaques. The main reason why plaques are so important is that we can harness and analyze the remains of the bacteriophage enzymes that carry out the lysing to understand the mutations of that phage, which was successful in lysing this one strain of bacteria.

Phages are great because they not only kill bacteria, but the resistance mechanisms the pathogens use fall flat when it comes to the phage’s ability to adapt in response to the bacteria’s resistome. Traditional phage therapy, however, has its own pitfalls. For one, phages are only able to infect very specific bacterial strains of species, making a phage useful for eliminating just one out of the potential thousands of pathogenic bacteria in a patient’s body. Furthermore, different phages utilize different mechanisms for lysis and have the chance to not even lyse or kill the cell at all.

Pillar 1: Revolutionizing Custom Phage Engineering.

Summary of Pillar 1:

The main disadvantage of phage therapy is the specificity of phage, meaning its ability to only target one specific strain of bacteria.

In this pillar, our primary goal is to engineer phage to increase its host range so that it can have a broader range of bacteria to infect.

BRED, or Bacteriophage Recombineering on Electroplated DNA is a highly effective homologous recombineering method that exploits the natural mechanism of phages: mutations. It works by inserting DNA into specific points of the host phage’s genome to cause mutations to the binding receptors. These mutations allow the phage to bind to multiple strains of bacteria.

Now we would test this phage’s efficacy of lysing the targeted bacteria. We can collect the plaque from the lysis of the bacteria, and using a Polymerase chain reaction, we can purify and analyze it to isolate the mutation. The successful mutation is analyzed against a naturally occurring mutation through polymerase chain reactions (PCR). The main goal is to identify exactly where the point mutations occurred, how effective they were and their feasibility for extraction.

All of this can ultimately revolutionize the process of discovering the most effective and viable mutation to increase the range of the phage.

Mutagenesis

Thanks to mutagenesis, we now don’t need to identify and engineer our desirable traits into phages, but we can now let the phages build these traits by applying constraints to them. Mutagenesis is the natural tendency of an organism to change its genetic code in response to stressors in the environment, and this is the same inherent system that bacteria and phages have used to fight each other for generations, and is also the same thing that antibiotics lack: the ability to adapt and change. Mutagenesis is especially key in increasing the phage’s host range.

Suppressing Bacterial Resistance through Phage Tail Fiber Mutagenesis

At Neuvae, we are trying to solve a phage’s host range, and this is quite the challenge because phages are tied to their host primarily because of their binding agents that are on the cell surface receptors — in order to recognize hosts and initiate an infection.

Since bacteria are easily able to mutate the sites that phages latch onto, phages undergo mutagenesis to make changes in their binding agents. As a result, most phages are composed of multiple, unrelated phages that collectively target a range of receptors and distribute the selective pressure away from any individual phage receptor.

This makes it harder for bacteria to defend against phage infections and is synonymous with the faster rate of mutation that bacteriophages are known for compared to bacteria. Despite this, the wide array of targets that these phages have naturally are not as effective at actually targeting and stopping a wide variety of pathogenic bacterial infections.

This prompts the question of whether we can change or alter a phage to bind to the desired host.

One study used targeted mutagenesis on the minor coat of a phage (essential for host recognition). The researchers used something similar for an antibody’s epitope of binding regions that they could engineer in vast amounts of host diversity for antigens.

The Phage Tail Fibers

The place of interest for using mutagenesis is the phage’s tail fibers. These tails give the phage their iconic shape and are capable of specific recognition of bacterial surfaces during the first step of viral infection.

The epitope recognition region of phages is directed by three regions called complementary determining regions on the tail proteins. Mutations in these regions can alter the target specificity of the antibody. Several phages like the T7 have tail fibers with similar regions which can be directly engineered through mutagenesis to increase their host range, and this has already been done quite successfully for treating E. Coli patients.

This prevents the need for expensive phage cocktails and allows for a single phage to become a library for multiple phage host targets. What this means is that for some phages like the T3 phage, you can increase the host range so that the phage can target naturally occurring phage-resistant bacteria mutations and prevent the onset of phage resistance.

Homologous Recombineering

Identifying these Host Range Determining Regions can be done through simple observation of the shape and homology. This prompts the idea of using homologous recombineering, which uses the structure and shape of a specific target to make genetic modifications to the organism. Through this, we can sequence mutations in these domains and identify their relation to host range using Phage display.

A T3 mutation is designed selectively for T3-resistant bacteria only, meaning that the T3 can naturally mutate its host range by being introduced to resistant bacteria. While this process is occurring, scientists found that introducing wild-type bacteria (a gene or phenotype that represents a bacterial species) will add pressure on the T3 to mutate and eliminate the bacteria in the process. This allows the T3 to now create new pathways that will attach to the bacteria with these wild types.

Think of it like sending innocent civilians in between a war. The army that sees the approaching waves of families will now have to change their objective to not just eliminating the enemy army, but also tending for the displaced families. Although grim, it paints a good picture of how selective pressure works in favour of the phage and can expedite the process of increasing a phage’s host range to not just inside of its species, but across a wide variety of species. Although unproven, this could potentially have the ability to make 1 phage a universal cure to many bacterial infections. That’s the goal we’re building up to, and we’re doing so with mutagenesis through homologous recombineering.

What’s great about recombineering is that it is time-efficient. The primary mutations that bacteria use to defend against foreign enemies are structural changes to their LPS (LPS mutations are changes in the receptor on the bacteria’s outside), and these mutations occur just after a few hours. Through random selection, unique mutations can be formed for the individual HRDRs and create more unique phage mutations to increase the host range.

This can be scaled up for mass production simply by turning the mutated sites into plasmids in order to selectively amplify the gene mutations that are important using PCR. You can now screen certain mutations and make the sequences accessible by contributing to a library and were found to yield productive phage mutants in receptor binding.

This method is also critical in mapping residues for receptor recognition and determining which amino-acid substitutions lead to host-range alterations, providing a rich tool for the structural biology of the receptor recognition.

Furthermore, homologous recombineering allows for mutagenesis to not just increase host range, but also possess different inhibiting mechanisms against bacterial resistance mechanisms. This also applies to enzymes like dispersion B in the T7 phage which can deliver biofilm-degradation through the enzyme. The engineered phage T7 expressed the DspB gene of Actinobacillus actinomycetemcomitans derived by the T7 φ10 promoter, which can be recognized by T7 RNA polymerase, therefore can significantly reduce the bacterial count in a single-species E. coli biofilm than the T7 phage control did. The same T7 was used to interfere with quorum sensing and inhibit biofilm formation, one of the primary resistance mechanisms.

This is where the idea of genetically modifying the target binding regions of a phage’s tail came to be, where we can just create 1 phage that could now expand its host range through its targeting mechanism. Now, you can formulate not just custom but scalable phages that can treat all strains of E.coli, or perhaps both E.coli and TB and so on. The only problem is that the most accurate methods out there are CRISPR-Cas9 and other genetic insertion/deletion methods which are just as costly and hard to scale as traditional phage cocktails.

Thankfully, we can exploit the natural mechanisms of phages and bacteria for our own benefit: mutations. Over years, bacteria have developed advanced genetic responses to invaders and threats that can trigger mutations through its resistome. It’s what makes antibiotic resistance such a challenge, but this is also why phages are able to fight back.

While antibiotics are static chemical compounds that do not evolve or change in response to AMR, phages are the biological competitor to bacteria and have, albeit different but effective, expansive genetic triggers that can allow them to mutate and circumvent the resistance mechanisms of bacteria.

This not only makes resistance less of a factor for phage therapy because the rate at which phages mutate and change is faster than that of the bacteria, but it also presents an opportunity for the phages to directly counteract the resistance mechanisms that are mutated (i.e. phages can better attack efflux pumps).

Through phage mutation, however, we can selectively force mutations in phages to attack multiple kinds of strains through homologous recombineering. The system comprises injecting DNA into specific points of the host’s genome or location. This can be used for altering the binding receptors of the phage to increase its range. However, traditional recombineering methods are ineffective at this because the phages cannot use selective markers such as drug-resistance genes for targeting the bacteria during viral lytic growth. Thanks to mutagenesis through homologous recombineering, this problem has been met but current systems are slow, unable to be used for all kinds of phages, and are not great at identifying useful mutations in the phages over several generations.

BRED System

This is where the BRED system comes in — BRED or Bacteriophage Recombineering of Electroporated DNA is a highly efficient recombineering system that primarily uses electroporated phage DNA. Electroporation is when you apply an electrical field to an organism and make its cell membrane more permeable for the insertion of DNA, drugs, and chemicals. In BRED, this means that mutants can easily be detected by PCR over the DNA (easily visible DNA) and it expedites the process significantly. BRED can be used to construct new, unmarked gene deletions, in-frame internal deletions, base substitutions, gene replacements with precision, and introduce gene tags for sequence identification. Learn More

Construction of a Gene Deletion using BRED

The benefit of this technique compared to previous iterations is that it can be applied to all mycobacteriophages and has been tested. Current sophisticated methods that have been used for mutant isolation and mapping are only applicable to commonly cited T4, T7, and lambda phages.

What BRED does differently to achieve a wider phage range is that it exploits a more general bacterial recombineering protein(s) gp60 and gp61. These are key in promoting resistance pressure and increasing the levels of recombineering potential for the individual phage.

Some techniques mediated by recombination proteins were used for E.coli using the lambda phage Red proteins, EXO AND BETA -> they then exploited Rac prophage for mutant construction, meaning that lytic growth is low and the role of antibiotic resistance pressure cannot be effectively used for mutant selection. Another key benefit is that thanks to the co-electroplating of both the phage DNA and targeting substrates, it becomes easier to engineer the substrate and its insertion, and when the mutations are then recovered, you can get a high proportion of copies that possess these mutations

Our pipeline’s first step involves exploiting the process of mutagenesis that phages are known for to engineer certain desirable traits such as specific resistance inhibitors, phage-derived enzymes to actively lyse the cell, and most importantly, increasing the range of the phage. This keeps the treatment competitive with antibiotics while making it far more effective at killing just the pathogenic bacteria above all else. We accomplish this through the BRED model that is able to synthetically engineer any large phage and was recently used to engineer a 4.3-kilobase pair Acinetobacter phage genome at a cost below $0.015 per base pair, hence enabling variant-engineered phage genomes to be obtained on demand and at a low price.

Sequencing the human genome right now through DNA sequencing tech costs approximately $0.07 for some of our mainstream therapeutics, and the potential for this process can tremendously expedite the production of far more effective phage therapy solutions at a price perhaps even cheaper than antibiotics. The BRED model also solves the acquisition of phages and functional analysis part of the pipeline, increasing the recovery rate from 0.01% to 33% for novel phages, further improving the efficiency of the purification and analysis part of the pipeline. What’s really awesome about this approach is that this cost only factors into the R&D part of the pipeline, as producing deliverable phage practices can be done through our host-reactor system for $8.90/500mL with a concentration of 4*1⁰¹⁰ phages/mL. This not only eliminates the need for novel discovery but can assist in further pushing the boundaries of the phage genome in ways that we might not even know of.

The recovery methods for these mutations are similar to most pipelines: the plaque that is formed through the phage replication and spread + lysis will be purified and analyzed against a wild-type (naturally occurring mutation) through PCR. When analyzing the selective mutant and wild type, you can quickly observe where the point mutations have occurred and depending on how easily the mutated types can be extracted, the more viable they will be for phage engineering. BRED can automatically expedite the process of identifying more useful and viable phage mutants as the process can recover 2–4x more pure mutations per plaque.

In fact, mutant-containing plaques can be recovered at an efficacy of 3.4–22.2%. After initial PCR screening of plaques, future purification is still needed to isolate homogenous phage mutants. Once the mutants are isolated, they will be then generated by infecting bacterial cells with the wild-type phage which would let the process to scale, following which extensive screening of recombinants will be required so that we can then identify the correct antibody for delivery using phage display.

Polymerase chain reaction (PCR) is a method used all over the world to multiply pieces of DNA from an initial strain. In our case, we can harness the power of PCR to screen and amplify the a screening and amplification matrix with the main goal of isolating the mutation from the phage and multiplying it so we can develop a large batch of phages that are successful in treating resistant bacteria.

The first step is to use PCR to isolate the mutation after harnessing and purifying the plaque. Plaque contains both wild-type phages and mutant phages, so both will be harnessed, however only the mutant phages will be put through the PCR multiplication process. Restriction enzymes can then be used to convert these mutations into specific point mutations, which is when the overall mutation can be translated down into the specific changes in each DNA nucleotide.

Now here is where the BRED system is used. The BRED system is especially important for recovering the pure mutations from the plaques because it can analyze more than double compared to traditional Flanking primer PCR methods. This is why we are using a combination of PCR and BRED, to maximize the benefits and bypass the drawbacks of each. This model can still be automated because using BRED for plaque screening is still very time-consuming due to the very specific and sensitive nature of BRED. Our system has also shown that mutants which cannot be derived are not viable for real applications, however, they can still act as a filter of sorts, reducing the time and effort required in analysis and sequencing. In terms of the success rate of the entire process, with current technology, mutant-containing phages can be recovered at an efficacy of anywhere between 3–22%, which will improve over time as the rate of technological developments increases. This percentage is for the recovery as well, future purification would still be needed to isolate the homogenous phage mutant.

Phage Display

The mutation would be developed through Phage Display for the mycobacteriophage and then used to construct gene deletions, replacements and heterologous gene insertions. This donor DNA contains the desired mutations flanked by homologous sequences of phage DNA to be engineered, which leads to homologous recombination occurring between the host phage genome and donor DNA. To make a deletion or small insertion, you will need to construct an approximately 200 bp double-stranded (dsDNA) substrate. For deletions, this should contain 100 base-pairs of homology upstream and downstream of the region to be deleted, making sure that deletion will be inframe, if necessary. For small insertions, this substrate should contain the sequence to be inserted, flanked by about 100 base pairs of homology on each end. To make a gene replacement mutant, you will need to construct a linear allelic exchange substrate (AES) that contains the sequence you wish to introduce, flanked by approximately 100 base pairs of sequence homologous to either end of the region to be replaced. mutants containing one or more point mutations can be generated using two synthetic complementary oligonucleotides. All of the substrates are co-transformed into electrocompetent recombineering cells with the phage DNA, and plaques are screened for the presence of mutation by PCR.

The problem with this system however is that it is very difficult in many cases to identify desirable bacteriophages that can actually lyse a bacteria when infected. We feel that phage-derived enzymes directly from virion-associated lysins, endolysin, and depolymerase can be used to lyse bacteria and can be used with any phage through intraperitoneal inoculation. The benefit of this is that they can lyse a wide range of species rather than strains. Several studies have exploited the fact that host range is linked to tail fiber composition for some phages. One scientist Yoichi genetically modified a T2 phage by swapping the long tail fiber genes (gp37 and gp38) with those from phage PP01, which specifically targets E. coli O157:H7. The exchange was done by homologous recombination between the genome of phage T2 and a plasmid carrying two regions of homology, flanking the gp37 and gp38 genes of PP01. As DNA synthesis, sequencing, and genome engineering tech will become more efficient, it will significantly expedite the possible host range.

We can also use phages for phagemids which encode plasmids to target certain resistance genes like for targeting the aph-3 kanamycin resistance gene that was packaged in the Staphylococcal phage ΦNM. Phasmids have been used to transfer foreign DNA across several bacterial species that helped express genes for protective antigens for a variety of pathogens.

Pillar 2: Enhanced Bacteriophage Targeting and Bacterial Lysis.

Summary of Pillar 2:

Bacteriophages can be used as a novel and efficient category of gene delivery vehicles for the introduction of various diagnostic and therapeutic cargoes to human cells. Using phages as delivery mechanisms is not toxic and poses no side effects.

These phages inject their genetic material into bacteria by recognizing the lipopolysaccharides, pili, peptidoglycans, proteins and teichoic acids comprising the cell walls and outer membranes of the bacteria.

In our solution, transducing particles are used as an antimicrobial agent that can go beyond lytic phage therapy. Particles are primarily used to transfer the inhibited resistance mechanism genes to other strains of bacteria. The assembly of transduction particles are comprised of the nucleotide cargo (inhibited resistance mechanisms), packages inside a viral protein.

Once we can engineer an ideal phage that can cross over several strains in a species for a general patient base, we then move on taking phage therapy to the next level. Studies have found that a common benefit of phage therapy is that the patient can see prolonged periods of tolerance to the bacterial strain of up to 32 weeks. This showcases that the active immune system has the potential to develop long-term resistance towards bacterial infections, something that was previously unable to be done. Imagine a vaccine that could treat a viral infection like COVID and prevent you from ever having it again. We can accomplish that through phage therapy assisted with transduction particles.

These particles are simply the necessary antigens required to trigger an immune response for a specific bacterial infection. The advent of phages has allowed for new technologies that can help identify the correct and most effective antigen for a given infection through phage display technology that aims to map phage-peptide relationships. Through a process called biopanning, the correct binding antigen is selected, washed and checked to see whether it can be used with the phage. This antigen is then packaged with a virion protein that has a natural affinity towards ACPs. The process goes as follows.

Once a phage would infect a bacteria, it can use the bacterial chromosome to produce transduction particles to make the bacteria’s resistance mechanisms weaker, assist in the bacteriophage lysis process, and code antigen production instructions for the dendritic cells. This can not only help in the production of new and general antigens but also automatically create an effective way to elicit a general immune response through the process of transducing these genetic instructions across different bacterial species. You can scaffold a transduction particle which would then allow the phages to inject these proteins and instructions into the bacteria (through the particles) and produce new, general antigens at the location of the bacteria. These antigens can be carried through the particles once more and attach to the dendritic cells. Now, instead of having to produce the antigen in a lab and transfer it, you can code the instructions onto the phage, use the phage on the site of the infection with presence in the bacteria, and use the transduction particles to create an antigen on-site across several different non-specific pathogenic bacteria without harming good bacteria thanks to the helper phages.

These particles are attached to the capsid of the phage to attract the active immune system. As the phages infect and lyse the bacteria, the transduction particles will be able to inhibit the resistance mechanisms the bacteria possess across multiple species as they are not considered foreign pathogens. This allows them to not just initiate an immune response thanks to the antigen expression on their surface but also dramatically expedite the therapy from a few days to just 2 hours. Once lycing is completed, the phages will be consumed through the innate immune system and phagocytosis whereas the antigen expression on the transduction particles will allow the Dendritic cells to develop a long-term memory of the genetic information the particles obtained during their spread thanks to the process of general genetic transduction. This also presents a unique opportunity to build a business model recurring around boosting your immunity every year or so, making the recurring nature of immunomodulation therapy more attractive for big pharma to adopt.

Pillar 3: Immunomodulation through Transduction Particles.

Summary of Pillar 3:

Traditionally, when a bacteriophage attacks a bacteria, an innate immune response is triggered and phagocytes are sent to kill the pathogenic bacteria. While this is effective in the short term, there is no memory of the bacteria.

This is the job of the adaptive immune system, which focuses on the development of antibodies against pathogens through the identification of unique antibodies, specifically the adaptive immune system. The innate response comprises phagocytes that engulf toxic materials such as lysed bacteria throughout an infection. This system naturally responds to any foreign pathogen without much direction. The adaptive immune system on the other hand is the system that uses antigens to produce antibodies. This directly encodes memory into the immune system and can reproduce these antigen-specific antibodies or peptides to attack the infection upon its onset. However, this is quite difficult to do with bacterial infections up until a recent paper showcased how patients that had phage therapy saw a 32 week increase in tolerance to the infection-causing bacteria. We want to exploit this while solving the fundamental problem. antibody production using a process called transduction. Our solution uses the transduction particles mentioned in the previous step to activate the production of antibodies against the bacteria. Triggering an adaptive immune response is the main alternative to antibiotics, the main method of killing the bacteria.

Antigens have a similar range problem like hosts, where 1 antigen can only be used for 1 specific bacterial strain. To circumvent this, We would then we’re fabricating something called a transduction particle to help trigger the immune system to remember the infection. Think of the particle like someone at costco. Initially, they would look for one specific sample and then end up eating a lot of these samples. The particle does something similar for each bacteria’s genetic chromosome through transduction. As the particle can hop around from each bacteria, it can use these gene samples to produce general antigens which attract the adaptive immune system to it. This is kind of like eating a lot of samples and then the manager has to come stop you. The manager is the body’s B cells, which can then utilize the random genetic samples the particle has taken + the antigens they’ve expressed to code antibodies that can later be used to defend against the same bacterial infection. The particle itself is also replicable through the phage’s infection mechanism, making the system scalable and cost-effective.

The first step is to use phage display technology to identify required antigens for the bacteria we are targeting through mapping phage-peptide relationships. Through a process called biopanning, the correct binding antigen is selected, washed and checked to see whether it can be used with the phage. After that, would engineer those specific antigens on the surface of the transduction particles.

The key to activating the adaptive immune system for a bacterial infection is the bacteriophages. Phages naturally have an affinity towards B lymphocytes, which will attract them to the location. Once the lymphocytes engulf the transduction particles, they will identify the antigens and activate the production of antibodies against those antigens, and therefore against the bacteria. This creates a general immune response which studies have found to keep resistance for up to a year. We can use this to form a proprietary business model surrounding taking an immune boost shot each year.

Scaling Plan

Our scaling plan begins with targeting the two deadliest bacterial infections:

MDR tuberculosis
Pneumonia

Our primary targets in R&D will be isolating TB, tapping into a 1.4 billion dollar market, and providing a better therapeutic for 10 million victims across the world. While our primary competitors are, like us, focused on providing a new drug for the multi-drug resistant TB strains, TB Alliance and Mylan are still in the antibiotic discovery field. We’re doing the same for Pneumonia and targeting younger outbreaks of macrolide-resistant strains in developing countries. 2.56 million people died from pneumonia in 2017, where almost a third of all victims were children younger than 5 years.

Sixty-four percent of the infections being treated in the ICU on any single day are pneumonia. What’s important to recognize here is that due to the ineffectiveness of antibiotics, most of the therapies used for these patients are second-phase treatments which add a greater strain on the healthcare system. Our goal with Neuvae is to take phage therapy from second-stage to first-stage by solving some of the most significant challenges from its adoption in the mainstream.

We’ll continue this trend by focusing on the WHO’s primary strains of concern, including

Enterobacteriaceae
Salmonellae
Staphylococcus Aureus
Pseudomonas Aeruginosa

Throughout each phase, we’ll primarily focus on answering some of the more practical solutions with production and engineering. This involves whether an inflammatory response will occur and how can we treat it (right now, looking into steroids), whether diagnosis tech for bacterial infections will continue to improve at its current rate, the toxicity of the lysed bacteria, the concentration of dose required, how to properly prescribe certain phage treatments, and who cannot take this therapy whether that be due to the potential side effects of our immunomodulation treatment or any other side effects that are unknown. We will also need to pass through each stage of preclinical and clinical trials.

From here, expanding the portfolio of phages we engineer and produce will play an important part in staying competitive with antibiotic manufacturers, but with advancements and funding for research in phage therapy and processes, we can save 25–30 billion dollars in global healthcare expenses, save up to 2.3 million lives each year, and live in a world where we might not even get sick from a chronic bacterial infection. It just might be possible, so let’s prove it and make it real.