Summary of Research Studies

A series of early studies were conducted at Los Alamos National Laboratory and in Company laboratories, as well as the 128-patient clinical trial (Patriquin, et al, Journal of Thoracic Oncology, September, 2015) that is summarized below. bioAffinity Technologies continues research to develop CyPath® as a diagnostic tool for early-stage cancer. New studies will be initiated to demonstrate increased sensitivity, specificity, positive predictive value and overall accuracy as a result of optimization completed after the clinical trial reported in Patriquin, et al.

Study Title and Description Study Subjects Principal Investigators & Institutions Results of Study
Study
Porphyrin’s localization in (abnormal) lung cells from uranium miners.
2 patients: One lung cancer patient and one person with COPD but no lung cancer D. Cole, PhD, Los Alamos National Laboratory (LANL); D. Moody, PhD, LANL; L. Ellinwood, MD, PhD, St. Mary’s Hospital; M. Klein, MD, St. Mary’s Hospital

Fluorescence was seen in every neoplastic (abnormal) cell identified in sputum samples, indicating high uptake of the bio-label.

The fluorescent intensity (brightness) of the cells was also greater in samples labeled with the porphyrin used by bioAffinity Technologies for cancer diagnosis and drug delivery as compared with samples labeled with other porphyrins.

Study
Testing the ability to diagnose lung cancer in different patients in a blind study with different types of lung cancers.
12 patients: Eight lung cancer / four non-cancer patients. D. Cole; D. Moody; L. Ellinwood; M. Klein

Sputum samples from cancer patients stained with the test showed the highest fluorescence with the exception of one patient who was thought not to have lung cancer but displayed high fluorescent cells in his sample. This person was re-evaluated and found to have lung cancer. With this finding, the study resulted in 100% accuracy.

Study
Testing in human squamous lung cancer cell line and ability of cells to be detected by automated imaging .
Lung Cancer Cell Lines D. Cole; D. Moody; L. Ellinwood; M. Klein

Squamous carcinoma (cancer) cells grown in culture stained with the bio-label could easily be detected with flow cytometry, an automated imaging system that measures the fluorescence of individual cells. Researchers determined that the auto fluorescence of control cells (cells not exposed to the bio-label) was negligible.

Study
Evaluation of the test on human small cell lung cancer and normal human lung epithelial cell lines
Cell Lines: 21 cancer samples and 4 non-cancer samples D. Cole; D. Moody; L. Ellinwood; M. Klein

100% of cancer samples contained high fluorescing cells as a result of high uptake of the bio-label. Concentration in normal and abnormal (non-cancer) lung cells was many times lower than cancer cells.

Study
Sputum Labeling for diagnosing lung cancer.
60 sputum samples; blinded study E. Lenkiewicz, MS, LungCheck, Inc.; R. W. Duke, PhD., LungCheck, Inc.

Abnormal cells (mild, moderate or severe dysplasia or cancerous) can be detected with the bio-label. Samples diagnosed as normal showed minimal or no uptake of the bio-label.

Internal Validation Research Trial 27 sputum samples: 15 lung cancer sputum samples and 12 sputum samples from patients without lung cancer T. Bauer II, MD, Helen F. Graham Cancer Center; D. Merrick, MD, Denver VA Medical Center; T. C. Kennedy, MD, University of Colorado Health Science Center; C. Dorian, Biomoda Sputum samples containing red fluorescing cells were considered positive for lung cancer. In this study, 15 of the 15 samples from lung cancer patients displayed red, fluorescing cells and 12 out of the 12 samples from normal patients reveals no red fluorescing cells, resulting in 100% accuracy in this dataset.
Clinical Trial
Early Detection of Lung Cancer with Meso Tetra (4-Carboxyphenyl) Porphine (TCPP)-Labeled Sputum, Journal of Thoracic Oncology, September, 2015
128 sputum samples from 26 diagnosed lung cancer patients and 102 military veterans at high risk for lung cancer L. Patriquin, MD, Radiology Assoc.; D. Merrick, MD, Denver VA Medical Center; D. Hill, MD, Waterbury Pulmonary Assoc.; R. Holcomb, PhD, Quintiles; V. Rebel MD, PhD, UTHSCSA; B. Karia, PhD, bioAffinity; G. Bennett, bioAffinity; T. Bauer II, MD, Meridian Health Systems The clinical study demonstrated 81% accuracy in detecting cancer from sputum samples collected from participants in two cohorts. The high-risk cohort included military veterans who had smoked at least 20 pack-years. The cancer cohort included patients diagnosed with lung cancer who provided sputum samples pre-treatment for the disease. A healthy cohort was added and analyzed resulting in 100% sensitivity and specificity. Study design limitations may have lowered accuracy in detecting cancer. For example, only a portion of the sputum sample was evaluated (approximately 30%). Optimization research following this trial supports improved accuracy and methods of automation for commercialization of a non-invasive, early lung cancer test.
Poster
Porphyrin uptake in lung cancer cells by dynamin-mediated endocytosis: a novel marker of dysregulated endocytosis in cancer. (American Society of Cell Biology Annual Meeting, 2015)
Research conducted on the biological Mechanism of Action (MOA) that causes cancer to preferentially take up CyPath D. Elzi, PhD, UTHSCSA; P. Fatland, UTHSCSA; B. Karia, PhD, bioAffinity; M. Iza, MD, UTHSCSA; A. Pertsemlidis, PhD, UTHSCSA; V. Rebel, MD, PhD, UTHSCSA The research reveals a biological process by which the bio-label is incorporated into lung cancer cells. The process, known as dynamin-mediated endocytotic pathway, is necessary for all cell growth, but is altered in cancer cells and results in increased uptake of the bio-label by cancer. Dysregulated endocytosis has been shown to be a hallmark of cancer.