ANIXA DIAGNOSTICS
LIQUID BIOPSY BASED ON FLOW CYTOMETRY AND AI TO DETECT THE EXISTENCE OF A SOLID TUMOR
Amit Kumar, Ph.D.
President and CEO
1---Subsidiary of ITUS Corporation
Technology uses the immune system as a signal amplifier
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IncreasingWBC Count
Solid TumorBacterial Infection
Healthy
Patients with solid tumors usually don’t have highly elevated WBCs
The frequency of certain Leukocytes sub-segments often change in tumor patients
Immuno-profiling can provide information regarding tumor status
WHITE BLOOD COUNT (WBC) INDICATES DISEASE
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• Malignancies do induce an immune response, though the increase in overall WBC count is modest• Tumors consist of cancer cells and immune cells• Research shows that high levels of a type of immune
cell called MDSC are found in malignant tumors• MDSC function is corrupted by the malignancy to
suppress general immune and T-cell functions.
Increased MDSC levels are also found in the circulation of cancer patients.
Myeloid Derived Suppressor Cell (MDSC)
MDSCS AND ONCOLOGYResearch supports the following conclusions
• Three phenotypically distinct MDSC populations
• E-MDSCs- Early
• M-MDSCs- Monocytic
• PMN-MDSCs- Poly-morphonuclear
• All MDSC are implicated in immune suppression
Published Literature
• Circulating MDSC levels correlate with Prognosis and Outcome• High MDSCs- poor outcome, poor response to therapy • Low MDSCs- better outcome, better response to therapy
• When Tumors are responding to therapy, circulating MDSC levels change• Monitoring of Tumor Response by MDSC analysis is being
conducted in multiple clinical trials• Other Lymphocytes also respond
• After a tumor has been resected, circulating MDSC levels change dramatically
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Hypothesis: Measuring MDSC levels could identify tumor-bearing patients
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BD LSR II High-speed Analyzer
How Do We Measure MDSCs? – Flow Cytometry
Increased MDSC Numbers in Peripheral Blood are Associated With Solid Tumors
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Angell et al. Thyroid 26(3) 2016.
Diaz-Montero et al. Cancer Immunol Immunother 58(1) 2009.
Zhang et al. PLoS ONE 8(2) 2013.
Dominguez et al. Clinical Cancer Research 23(12) 2017.
Thyroid BreastColorectal
Advanced Solid Tumors Lung
Chen et al. Oncology Letters 14 2017.
MDSCs can be measured in peripheral blood using flow cytometry – a process
known as immunophenotyping
Cancer Stage
MDS
C Co
unt
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ARTIFICIAL INTELLIGENCE ANALYSIS
Training Calls
Neural NetworkNormal Patient Samples
Cancer Patient Samples
Patient Sample
Malignancyor
Normal
Breast CancerPredicted
Naïve Testing All Samplesn = 56 n = 144
TB nTB TB nTB
Measured
TB 24 2 58 6nTB 4 26 11 69
Sens. (%) 92.31 90.62Spec. (%) 86.67 86.25Acc. (%) 89.29 88.19PPV (%) 85.71 84.06NPV (%) 92.86 92AUROC(95% CI)
0.8987(0.8047 to 0.9927)
0.9209(0.8705 to 0.9713)
ROC analysis: NN vs Manual Gating Vs Mammography/Ultrasound
All Naïve Samples were Early Stage (Stage 1 and 2)
Prostate CancerPredicted
Naïve Testing All Samplesn = 36 n = 146
TB nTB TB nTB
Measured
TB 16 2 68 5nTB 1 17 1 72
Sens. (%) 88.89 93.15Spec. (%) 94.44 98.63Acc. (%) 91.67 95.89PPV (%) 94.12 98.55NPV (%) 89.47 93.51AUROC(95% CI)
0.929(0.8418 to 1)
0.9797(0.9592 to 1)
ROC analysis: NN vs Manual Gating vs PSA Test
N = 73Predicted
Manual Gating Neural NetworkProstate Healthy Prostate Healthy
Measured
Prostate 16 19 33 3Healthy 10 28 3 34
Sens. (%) 45.71 91.67Spec. (%) 74.36 91.89Acc. (%) 60.81 91.78PPV (%) 61.54 91.67NPV (%) 60.42 91.89AUROC(95% CI)
0.597(0.4658 to 0.7282)
0.9354(0.8723 to 0.9985)
*Manual gating assumed a 75% specificity as the cutoff
Prostate Cancer Study:Memorial Sloan Kettering Data
Key Conclusions• Artificial Intelligence (AI) better than current methods used by scientists or physicians to make calls
• May work for all cancers
• To date, identified in 20 different types- May work for all tumor types
• Multiple samples:
• Single sample:
• Cost to run test is a few dollars12
Breast Prostate Lung Colon
Pancreatic Melanoma Ovarian Liver
Bladder Cervical Endometrial Thyroid
Gastric Head and Neck Testicular
Osteosarcoma (bone cancer)
Leiomyosarcoma (cancer of the soft tissue)
Liposarcoma (cancer of the connective tissue)
Vulvar
Appendiceal