Precision Pathology

Please visit our Physician Portal regarding our CyPath^® Lung Test Kit, featuring important items like How the CyPath^® Lung Test Works, How to order CyPath^® Lung, and Instructions for you and your patients:

CyPath^® Lung Physician Portal

Please visit our Patient Portal regarding our CyPath™ Lung Test Kit, featuring important information about CyPath™ Lung, and how to collect and return your sputum sample to the laboratory:

CyPath^® Lung Patient Portal

Detection of cancer at its earlier stages is key to reducing cancer mortality. At an early stage, cancer is more likely to still be localized, not spread to other parts of the body yet, which makes treatment easier and more successful. People at high risk for lung cancer are recommended for annual screening with low dose computed tomography (LDCT) to find cancer at earlier stages when it can be successfully treated. The National Lung Cancer Screening Trial1 (NLCST) of more than 53,000 high-risk patients showed LDCT screening could lower mortality rates by 20% as compared to screening with traditional X-ray. It is very important for people at high risk for lung cancer to be screened annually for the disease. Early detection of lung cancer is key to increasing the survival rate of patients.

While widespread use of LDCT will help lower the death rate from lung cancer, the NLCST found that LDCT screening has a low positive predictive value of only 3.8%. That means that for every 100 people who were diagnosed by LDCT as positive for lung cancer, less than four of them actually had the disease. The low positive predictive value can lead to more expensive and riskier procedures to diagnose lung cancer. People may decide not to be screened because of the anxiety, cost and risk of a positive LDCT result that turns out to be incorrect. That is a mistake. Screening for lung cancer is essential if we want to diagnose early-stage cancer and save lives.

A patient-friendly, accurate and cost effective test like CyPath ^® Lung offers significant benefits. The use of CyPath^® Lung after a positive LDCT scan can increase the positive predictive value of screening and result in lowering the number of people who would otherwise have to undergo unnecessary and invasive procedures and provide physicians with greater confidence when follow-up is necessary.

Current diagnostic methods for lung cancer include PET scans, bronchoscopy and tissue biopsy. The table below compares the sensitivity and specificity of CyPath^® Lung with tests and procedures currently used to diagnose lung cancer. CyPath^® Lung compares favorably to these more invasive and expensive tests.

The table below compares the sensitivity and specificity of CyPath® Lung with tests and procedures currently used to diagnose lung cancer. CyPath® Lung compares favorably to these more invasive and expensive procedures. A test validation trial comparing people at high risk for lung cancer to patients with the disease resulted in CyPath® Lung specificity of 88% and sensitivity of 82%, similar to far more invasive procedures currently used to diagnose lung cancer. CyPath® Lung performed even better, with 92% sensitivity and 87% specificity, in the group of cancer and cancer-free high-risk participants who had small nodules less than 20 mm in diameter or displayed no nodules. The high sensitivity and specificity of CyPath® Lung for smaller nodules is important because nodules between 6 mm and 20 mm can pose more difficulty with diagnosis. CyPath® Lung can provide more certainty to physicians and their patients.

 

*Specificity is a measure of how many times the test correctly indicates that a person who does not have the disease is negative.

**Sensitivity is a measure of how many times the test correctly indicates that a person who does have the disease is positive.

 

Diagnostic Method	Specificity*	Sensitivity**
CyPath Lung®	88%	82%
CyPath Lung® (Nodules < 20 mm)	87%	92%
Low Dose CT (LDCT)1	73.4%	93.8%
FDG PET Scan2	82%	87%
Bronchoscopy3	47%	88%
Fine Needle Biopsy4	75.4%	90.4%
Core Needle Biopsy4	88.6%	89.1%

CyPath® Lung interrogates the lung micro-environment by analyzing sputum using flow cytometry. The test acquires information about sputum using a flow cytometer that individually scans and characterizes tens of millions of cells in minutes. CyPath® Lung labels a person’s sputum sample with fluorescent antibodies that uniquely identify different cell types and a fluorescent porphyrin called TCPP that preferentially binds to cancer cells and cancer-related cells. TCPP fluoresces a bright red color when attached to cancer cells and cancer-associated cells that is visible when the sample is run through a flow cytometer. CyPath® Lung uses automated analysis that looks for pre-set parameters that are predictive of cancer, one of which is the presence of cells that have taken up greater amounts of the TCPP porphyrin label.

A diagnostic should be tested in the population in which it will be used. In the case of lung cancer, the relevant population is people at high risk for the disease. As LDCT screening shows, it is difficult to tell the difference between someone with cancer and someone at high risk without cancer. Smoking severely damages the lungs, leading to respiratory conditions that confound analysis and can produce nodules that look like cancer on scans.

Many clinical studies of diagnostics on the market or in development compare cancer patients with healthy individuals, resulting in seemingly higher accuracy but lacking clinical relevance. The pathology of a healthy lung is very different from one with cancer, in contrast to the lungs of a high-risk individual without cancer who has endured the insult of smoking in comparison to the lungs of patients with lung cancer. The CyPath® Lung trial compared these two very similar but difficult to distinguish cohorts – those at high risk with and those without cancer – to provide meaningful results for clinical use.

The test validation of CyPath® Lung rests on a solid scientific foundation. Porphyrins have been known for decades to specifically concentrate in many types of cancer cells and cancer-associated cells such as macrophages that are cells that clean debris from the tissues in the body. In addition, certain porphyrins are naturally fluorescent, with a distinct photon emission profile. Early research conducted at Los Alamos National Laboratory showed 100% accuracy in distinguishing lung cancer patients from healthy individuals by labeling sputum with the synthetic porphyrin TCPP. A later sputum cytology study published in 2015 showed that certain cells in sputum from lung cancer patients labeled by the porphyrin TCPP fluoresce a bright red color that can be measured under a microscope. In this trial, the results of which were published in 2015, study participants were classified into cancer versus high-risk cohorts with 81% accuracy. Building on this scientific foundation, the current CyPath® Lung test still uses porphyrins but adds other markers to the analysis. In addition, the test is now performed on a flow cytometry platform which makes it possible to quickly analyze millions of cells in a single sputum sample.

CyPath® Lung is the first non-invasive, accurate and well-balanced test for lung cancer, meaning that our test has high specificity and high sensitivity, compared to other tests that claim high sensitivity and very low specificity, or vice versa.

CyPath® Lung investigates the lung micro-environment by analyzing sputum, using antibody staining of cells similar to the flow cytometry analysis of blood for lymphomas and other blood-related cancers. CyPath® Lung uniquely incorporates the porphyrin TCPP to identify cancer and cancer-related populations in sputum. Test results that are provided to the physician ordering CyPath® Lung are generated using automated analysis that is completed in minutes.

Exfoliated cells in sputum have long been of interest for lung cancer diagnosis. Traditional slide-based sputum cytology, which looks at only a fraction of the entire sputum sample, has been shown to detect lung cancer in 60% to 82% of cases⁶.

Sputum bathes the lungs, coming into direct contact with a possible tumor that sheds cancerous and cancer-related cells, making sputum an excellent sample for cancer detection.

Sputum can be collected easily at home, either spontaneously or with assistance from a non-invasive, hand-held airway clearing device that helps people expel the sample into a collection cup.

A physician orders CyPath® Lung and gives the patient a sputum collection kit to take home. The kit includes one test collection cup; a pre-addressed, overnight envelope for returning the sample; a patient instruction booklet; and a small cold pack to be placed in the freezer during the three-day collection period. Patients collect a sputum sample at home every day for three consecutive days, expelling sputum and storing the three-day sample in the same collection cup. Sputum is expelled and collected by coughing into the collection cup. The patient stores the collection cup in a refrigerator between collections.

On the third day of collection, the patient places the collection cup containing the sputum sample back into the kit box for return by overnight mail to the laboratory. The patient adds the frozen cold pack to the kit box that contains his or her sputum sample prior to sending it to the laboratory for analysis.

bioAffinity Technologies plans to develop additional cancer diagnostics using its flow cytometry platform. The Company also plans to develop tests related to the health of the lung, including diagnostics for asthma and COPD. Additional research and development are required to bring these tests to market.

Several clinical trials are planned including the following:

• CyPath^® Lung Pivotal Clinical Trial is a prospective, multi-site trial enrolling approximately 2,000 high-risk patients at 20 collection sites over a two-year period that will establish **sensitivity and *specificity in distinguishing high-risk individuals with cancer from those who are cancer-free but have a suspicious lung nodule revealed by LDCT screening.
• Pivotal Clinical Trial Combining LDCT with CyPath^® Lung is a prospective, multi-site trial enrolling approximately 2,000 high-risk patients over a two-year period to establish **sensitivity and *specificity of combined testing for participants with suspicious nodules revealed by LDCT who are subsequently tested by CyPath^® Lung.
• Clinical Utility Trial of CyPath^® Lung is a prospective, multi-site trial to determine how physicians currently manage patients with suspicious lung nodules and how the results of combined testing including CyPath^® Lung will change clinical decision-making.

Most liquid biopsies are used after a tissue biopsy has confirmed a diagnosis to identify specific gene mutations in the tumor cells. This information helps inform therapeutic choices by giving physicians the opportunity to select drugs that target the cellular pathways that are affected by the gene mutations. CyPath® Lung detects lung cancer in high-risk individuals prior to a tissue biopsy or other confirmatory tests that establish the person has lung cancer.

Some liquid biopsies seek to find cancer earlier. So far, diagnostic liquid biopsies for early lung cancer show high sensitivity and very low specificity, or vice versa. Low sensitivity means a test misses the majority of cancers for which it is testing, leading to high false negatives. Low specificity means the test misses the majority of people who do not have cancer, instead calling them positive, leading to many unnecessary follow-up tests. CyPath® Lung is different, showing both high specificity and sensitivity in testing for lung cancer.

CyPath® Lung analyzes the lung micro-environment by looking for pre-set parameters indicative of having lung cancer. Flow cytometry, the platform used by CyPath® Lung for analysis, profiles cell populations, i.e., whole cells, to detect disease. Liquid biopsies most often analyze blood samples. Although some identify whole cells as well (so-called circulating tumor cells), most liquid biopsies look for genetic fragments of cancer cells in blood.

Sputum is much more concentrated with cancer-related cells and cancer-related material than blood, providing a better sample to analyze for lung cancer.

Several companies are developing “pan-cancer” blood tests (liquid biopsies), meaning that one blood draw could be used to detect one or more of 50 cancers. This approach limits the search for specific mutations, which is especially problematic for lung cancer since lung cancer is known to be associated with many different types of mutations.

Sputum travels well. A three-day sample overnighted to the laboratory exhibits high viability, with on average more than 60% of cells arriving alive for processing. In comparison, blood can be especially vulnerable to handling and processing problems. DNA, RNA and proteins tend to degrade quickly when processed inappropriately. Degradation of test targets is a particular concern when the targets are rare, as is the case with early-stage cancers.

CyPath® Lung is billed as basic flow cytometry test.

For more information regarding billing, click here.

¹ National Lung Screening Trial Research Team, Church TR, Black WC, Aberle DR, Berg CD, Clingan KL, et al. Results of initial low dose computed tomographic screening for lung cancer. N Engl J Med. 2013 May 23;368(21):1980-91
² Deppen et al., Accuracy of FDG-PET to diagnose lung cancer in areas with infectious lung disease: A meta-analysis, JAMA, 2014
³ Silvestri et al. A Bronchial Genomic Classifier for the Diagnostic Evaluation of Lung Cancer, New England Journal of Medicine, 2015
⁴ Yao et al, Fine-needle aspiration biopsy versus core-needle biopsy in diagnosing lung cancer: a systemic review, Current Oncology, 2012
⁵ Patriquin L, Merrick DT, Hill D, Holcomb RG, Lemieux ME, Bennett G, et al. Early Detection of Lung Cancer with Meso Tetra (4-Carboxyphenyl) Porphyrin-Labeled Sputum. J Thorac Oncol. 2015 Sep 1;10(9):1311–8
⁶ Ng AB, Horak GC. Factors significant in the diagnostic accuracy of lung cytology in bronchial washing and sputum samples. II. Sputum samples. Acta Cytol. 1983