High intensity focused ultrasound and methods of performing non-invasive biopsies using same

Abstract

The present disclosure provides methods of performing a liquid molecular biopsy on a subject using high intensity focused ultrasound (HIFU) energy. The present disclosure also provides methods of diagnosing a disease or a risk of a disease, such as a cancer, in a subject. The present disclosure further provides methods of treating one or more tissue masses (e.g., nodules, tumors, cysts, lesions, cells of unknown significance etc.) in a subject. In some embodiments, the methods are non-invasive or minimally invasive.

1 - 14 . (canceled) 
     
     
         15 . A method of treating a target tissue mass in a subject, the method comprising:
 optionally determining a baseline concentration of a biomarker in a fluid sample of the subject;   performing a procedure on the target tissue mass of the subject;   applying high intensity focused ultrasound (HIFU) energy to the target tissue mass;   determining a concentration of the biomarker in a fluid sample of the subject after applying HIFU energy to the target tissue mass; and   repeating the performing, applying, and determining until a concentration of the biomarker in the fluid sample falls below a threshold value.   
     
     
         16 . The method of  claim 15  wherein a period of time between applying HIFU energy to the target tissue mass and determining the concentration of the predetermined biomarker is no more than about 2 hours. 
     
     
         17 . The method of  claim 15  wherein a period of time between applying HIFU energy to the target tissue mass and determining the concentration of the predetermined biomarker is no more than about 1 hour. 
     
     
         18 . The method of  claim 15  wherein a period of time between applying HIFU to the target tissue mass and determining the concentration of the predetermined biomarker is no more than about 20 minutes. 
     
     
         19 . A method of treating two or more tissue masses in a subject, the method comprising:
 (a) optionally determining a baseline concentration of a first marker and/or second marker in fluid of the subject;   (b) performing a first procedure on a first tissue mass of the subject;   (c) applying high intensity focused ultrasound (HIFU) energy to the first tissue mass to cause a first marker to release from the first tissue mass;   (d) thereafter, determining a concentration of the first marker in a first fluid sample of the subject;   (e) after a first period of time, performing a second procedure on a second tissue mass of the subject;   (f) applying HIFU energy to the second tissue mass to cause a second marker to release from the second tissue mass;   (g) thereafter, determining a concentration of the second marker in a second fluid sample of the subject; and   (h) after a second period of time, repeating:
 (1) steps (b) to (d) if the concentration of the first marker exceeds a first threshold value; 
 (2) steps (e) to (g) if the concentration of the second marker exceeds a second threshold value; or 
 (3) steps (b) to (g) if the concentration of the first marker exceeds the first threshold value and the concentration of the second marker exceeds the second threshold value. 
   
     
     
         20 . The method of  claim 19  wherein the first threshold value is significantly greater than the baseline concentration of the first marker in fluid of the subject determined before step (c), optionally wherein the threshold value is about 2 times greater than the baseline concentration of the first marker. 
     
     
         21 . The method of  claim 19  wherein the second threshold value is significantly greater than the baseline concentration of the second marker in fluid of the subject determined before step (f), optionally wherein the threshold value is about 2 times greater than the baseline concentration of the second marker. 
     
     
         22 . The method of  claim 19  wherein applying HIFU energy to the first mass and applying HIFU energy to the second tissue mass induces boiling histotripsy or cavitation. 
     
     
         23 . The method of  claim 19  wherein the fluid sample is obtained from the subject while applying HIFU energy to the subject. 
     
     
         24 . A method of inducing release of a marker from target tissue of a subject, the method comprising non-invasively applying high intensity focused ultrasound (HIFU) energy to the target tissue. 
     
     
         25 . The method of  claim 24  wherein the target tissue is a tumor and the marker is an miRNA. 
     
     
         26 . The method of  claim 24  wherein a maximum amount of the marker is released from the target tissue in no more than about 5 minutes after application of the HIFU energy. 
     
     
         27 . (canceled) 
     
     
         28 . The method of  claim 26  wherein the marker is released into a fluid of a subject, and the amount of the marker released from the target tissue is determined by determining a concentration of the marker in a fluid of the subject, wherein the fluid is optionally selected from the group consisting of blood, a blood fraction, a blood derivative, a fluid fraction of blood, a non-fluid fraction of blood, whole blood, blood cells, microvesicles, cryoprecipitate, plasma, serum, subcellular vesicles, lymph fluid, ascites, urine, cerebrospinal fluid, seminal fluid, breast milk, breast secretions, breast aspirates, and feces. 
     
     
         29 . The method of  claim 28  wherein the fluid is blood, a blood fraction, or a blood derivative. 
     
     
         30 . The method of  claim 26  wherein the maximum amount of the marker is significantly greater than the baseline amount of the marker in the fluid before application of the HIFU energy, optionally at least about 2 times greater than the baseline amount. 
     
     
         31 . The method of  claim 19  wherein the first marker and the second marker are the same. 
     
     
         32 . The method of  claim 15  wherein the marker comprises any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6. 
     
     
         33 . The method of  claim 19  wherein the marker comprises any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6. 
     
     
         34 . The method of  claim 24  wherein the marker comprises any one of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6. 
     
     
         35 - 37 . (canceled) 
     
     
         38 . The method of  claim 15  wherein the subject is human. 
     
     
         39 . The method of  claim 15  wherein the subject is a non-human animal. 
     
     
         40 . The method of  claim 19  wherein the subject is human. 
     
     
         41 . The method of  claim 19  wherein the subject is a non-human animal. 
     
     
         42 . The method of  claim 24  wherein the subject is human. 
     
     
         43 . The method of  claim 24  wherein the subject is a non-human animal.