Endovascular bioelectronic device and method for detection of circulating biomarkers

Abstract

An exemplary implantable active bioelectronics stent can be provided. For example, the stent can comprise a mechanically flexible package, an active integrated circuit (IC) chip associated with the flexible package; and one or more electrodes configured to sense biomarkers in a bloodstream and associated with the active IC chip. The stent can use wireless powering and communication via electromagnetic or ultrasound energy.

Description

93597 / 7621ENDOVASCULAR BIOELECTRONIC DEVICE FOR DETECTION OF CIRCULATING BIOMARKERS AND METHOD FOR THE USE OF THE ENDOVASCULAR BIOELECTRONIC DEVICECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit of U.S. Provisional Application No. 63 / 742,261, filed January 6, 2025 and U.S. Provisional Application No. 63 / 767,707, filed March 6, 2025, the contents of which are hereby incorporated by reference.FIELD OF THE DISCLOSURE

[0002] The present disclosure relates generally to wearable devices, and more particularly to wearable device(s) for the delivery' of focused light into body and methods for the use of the wearable device(s).BACKGROUND INFORMATION

[0003] Death from metastatic cancer remains one of the leading causes of mortality from cancers; hence, methods to detect cancer early are urgently required [see. e.g., Ref. 1], In the treatment of cancer, radically new approaches are needed for an early detection, monitoring treatment response, and assessing the risk of metastasis or recurrence. Biopsies and molecular profiling of tumor tissues are invasive, provide limited information, and are often impractical after tumor removal. As an alternative, blood-based biomarkers, known as liquid biopsies, show promise because they can be monitored through a simple blood draw, which is less invasive and allows for more frequent assessments throughout treatment [see, e.g., Ref. 2], Currently, blood-based cancer biomarkers are divided into non-cell-based markers, such as cell-free DNA (cfDNA), proteins, and exosomes, and cell-based markers [see, e.g.. Ref. 3], Protein biomarkers have been used for monitoring disease progression, although detection is not always accurate in every patient [see, e.g.. Ref. 4], Other markers, such as cfDNA, provide only partial insights into the disease, as cfDNA gives limited genomic data without accompanying protein information, and protein markers like CEA lack complementary tumor-specific genetic data [see, e.g.. Ref. 3],

[0004] The process of metastasis involves the tumor cells leaving the primary site and landing and establishing at distant sites [see, e.g., Refs. 5 and 6], Circulating tumor cells (CTCs) shed directly into the bloodstream, serve as valuable indicators of metastatic4935-6609-1653v.lpotential [see, e.g., Ref. 7], Liquid biopsies based on blood testing have been believed to be an approach for an early detection based on the detection of the CTCs [see, e.g., Ref. 8], As such, CTCs have been some of the most extensively studied cell-based cancer biomarkers [see, e.g.. Refs. 9-12], However, CTCs are rare in peripheral blood samples, typically less than five CTCs per 7.5 mL in high-burden cases, making them challenging to isolate and evaluate.

[0005] Recent research from many groups has unveiled a new category of CTCs known as circulating hybrid cells (CHCs) or tumor hybrid cells (THC) [see, e.g., Refs. 13- 28], These entities express both epithelial and immune cell markers, and their presence was demonstrated in the tumor biopsies of patients with colon and breast cancers [see, e.g.. Ref.29], The CHC are usually characterized as being large and usually polyploid; as such they have been termed polyploid giant cancer cells (PGCC) or cancer-associated macrophagelike cells (CAML). CHCs. as a recently identified type of CTC. express both immune and tumor proteins and have shown promise as a novel biomarker for cancer [see, e.g., Refs. 19, 20 and 29-31], Likely arising from the fusion of immune cells and tumor cells, CHCs display unique properties, such as enhanced migratory abilities and increased potential for metastasis [see. e.g., Ref. 20], These cells are the dominant tumor-related cell population in peripheral blood across many cancer types, and their levels in the blood of cancer patients correlate with disease stage and prognosis [see, e.g., Refs. 19, 20 and 31],

[0006] While ex vivo capture devices have been developed, most are limited to small blood volumes (1-50 mL) due to safety concerns [see, e.g., Ref. 32], The use of these devices likely restricts the absolute number of CTCs obtained and captures only those present at a specific moment, overlooking temporal fluctuations in release [see, e.g., Ref.29], Several FDA-approved and commercial platforms for the detection of CTCs include CellSearch [see, e.g., Ref. 33], which employs immunomagnetic isolation; Vortex [see, e.g., Ref. 34], which employs size-based microfluidic separation; and DEP Array [see, e.g., Ref.35], which uses dielectrophoresis to trap cells. It is believed that none of these current technologies address one of the biggest challenges with these liquid biopsy approaches, which is the scarcity7of CTCs.

[0007] Extending the volume interrogated over longer time periods is important to enhancing CTC (and CHC) utility as biomarkers by increasing the statistical confidence. Cytapheresis products, often collected for hematopoietic stem cell transplantation therapy, can provide a substantial increase in detectable CTCs compared to single blood draws [see,4935-6609-1653v.le.g., Ref. 36], Yet, standard cytapheresis primarily consists of concentrated peripheral blood mononuclear cells, requiring additional screening steps for CTC identification. It is believed that the only in vivo approach that has been used is one that trap cells labelled with magnetic beads with the use of a magnetic wire that captures the cells through a catheter for removal [see, e.g., Ref. 37], Other work likely uses an indwelling intravascular aphaeretic device to isolate CTC [see, e.g., Ref. 38], Both require cell remove and rely on pre-injected magnetic particles for labelling.

[0008] Thus, there is a need to address and / or improve various issues and / or deficiencies which exist in the previous devices, systems, and processes.SUMMARY

[0009] An implantable active bioelectronics stent, comprising:

[0010] a mechanically flexible package;

[0011] an active integrated circuit (IC) chip associated with the flexible package; and

[0012] one or more electrodes configured to sense biomarkers in a bloodstream and associated with the active IC chip, wherein the stent is powered by wireless powering and communicated with via electromagnetic or ultrasound energy

[0013] A device comprising:

[0014] an implantable active bioelectronics stent which comprises:

[0015] a mechanically flexible package;

[0016] an active integrated circuit (IC) chip associated with the flexible package; and

[0017] one or more electrodes configured to sense biomarkers in a bloodstream and associated with the active IC chip, wherein the device uses wireless powering and communication via electromagnetic or ultrasound energy.

[0018] A kit comprising a device described herein, and an external wearable device for placing in close proximity with said device and communicating with said device.

[0019] A method of diagnosing metastatic potential in a subject comprising detecting a circulating tumor cell (CTC) in a bloodstream of a subject using a device described herein by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device, wherein presence of a CTC in the bloodstream is indicated by differential EIS indicative of a CTC and indicates metastatic potential, and wherein absence of detection of a CTC in the bloodstream does not indicate metastatic potential.4935-6609-1653v.l

[0020] A method of reducing the likelihood of metastasis in a subject comprising detecting a circulating tumor cell (CTC) in a bloodstream of a subject using a device described herein by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device, wherein presence of a CTC in the bloodstream is indicated by differential EIS indicative of a CTC and indicates likelihood of metastasis, and wherein when the CTC is detected electroporating the CTC via the device.

[0021] A method of treating a hematological cancer in a subject comprising detecting a hematological cancer cell in a bloodstream of a subject using a device described herein by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device, wherein presence of a hematological cancer in the bloodstream is indicated by differential EIS indicative of a hematological cancer cell and, wherein when the hematological cancer cell is detected, electroporating said cell via the device.

[0022] A method of destroying a predefined cell type or type of plurality of cells in a subject comprising detecting a predefined cell type or type of plurality of cells in a bloodstream of a subject using a device described herein by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device, wherein presence of a predefined cell type or type of plurality of cells is indicated by differential EIS indicative of a predefined cell type or type of plurality’ of cells, respectively, and wherein when the predefined cell type or type of plurality of cells is detected, electroporating said predefined cell type or type of plurality of cells via the device.

[0023] A non-limiting exemplary’ implantable active bioelectronics stent according to an exemplary embodiment of the present disclosure is be provided. For example, the stent can comprise a mechanically7flexible package, an active integrated circuit (IC) chip associated with the flexible package; and one or more electrodes configured to sense biomarkers in a bloodstream and associated with the active IC chip. The stent can use wireless powering and communication via electromagnetic or ultrasound energy. A device that can include such stent is also be provided.

[0024] For example, such one or more electrodes are used to (i) perform an electrochemical impedance spectroscopy of cells in the bloodstream and / or (ii) electroporate cells in the bloodstream.

[0025] The powering of the device can be through a near-field inductive coupling. The communication to and / or from the stent can occur over one or more impulse-radio, ultra-wide-band communications links. The stent can be mounted on a bioresorbable scaffold.4935-6609-1653v.lThe bioresorbable scaffold can be composed of poly-L-lactide. The active IC chip can be mechanically thinned so as to be mechanically flexible. The flexible package can integrate therein or thereon a metallic coil. The active IC chip can include an antenna thereon or therein. In embodiments, a external wearable device is placed over a location of the stent.

[0026] These and other objects, features, and advantages of the exemplary embodiments of the present disclosure will become apparent upon reading the following detailed description of the exemplary embodiments of the present disclosure, when taken in conjunction with the appended paragraphs.BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Further objects, features and advantages of the present disclosure will become apparent from the following detailed description taken in conjunction with the accompanying Figures showing illustrative embodiments of the present disclosure, in which:

[0028] Fig. 1 is a diagram of an exemplary “Stentinel” system, according to exemplary embodiments of the present disclosure;

[0029] Fig. 2A is an illustration of a polyimide package design for the exemplary Stentinel system for a 4-mm-diameter vessel with a vessel-wall-facing side containing a 15mm x E5 mm ASIC of the exemplary Stentinel system and the wireless powering coil, according to exemplary embodiments of the present disclosure;

[0030] Fig. 2B is an illustration of the polyimide package design for the exemplary Stentinel system for a 4-mm-diameter vessel with an interior-side of the package of the exemplary Stentinel system containing the electrodes of the EIS modules, according to exemplary embodiments of the present disclosure;

[0031] Fig. 3 is an exemplary' illustration of an exemplary differential EIS measurement of cells in a half-bridge topology from the three-electrode detection module, according to exemplary embodiments of the present disclosure;

[0032] Fig. 4 is a set of schematic diagrams of exemplary designs of the exemplary Stentinel system, according to exemplar}' embodiments of the present disclosure;

[0033] Fig. 5A is a graph of power transfer efficiency vs. distance as function of coupling coefficient to provide an analysis of the wireless powering of the Stentinel, according to exemplar}' embodiments of the present disclosure;4935-6609-1653v.l

[0034] Fig. 5B is an exemplary illustration of a simulated S21 and power transfer efficiency for the relay station coil coupled to the on-package coil on the exemplary Stentinel system, according to exemplar^' embodiments of the present disclosure;

[0035] Fig. 6 is a set of graphs, circuit design and tables providing exemplary electrochemical impedance spectroscopy from 20 pm x 20 pm TiN electrodes, according to exemplary embodiments of the present disclosure;

[0036] Fig. 7 is an illustration of an exemplary representation of the relay station that has two modules, according to exemplary embodiments of the present disclosure, and also clotan exemplary representation of the relay station that consolidates all components into a single module, according to exemplar}' embodiments of the present disclosure;

[0037] Fig. 8 is a diagram providing an exemplary' software infrastructure of the exemplary' Stentinel system, according to exemplary embodiments of the present disclosure.

[0038] Figs. 9A-9B show an exemplary custom stent design. (A) Flattened view of the stent design (dark grey) with the unrolled polyimide containing the electrodes (light grey). (B) 3-D view of the stent with the large open cells (highlighted blue) aligned with the arrangement of the electrodes (semi-transparent);

[0039] Figs. 10A-10B show an exemplary stent structure (A) that incorporates electrodes and wiring onto the stent struts, including an exemplary detail as shown in (B);

[0040] Figs. 11A-11O show an exemplary7Polyimide Thin-Film Electrode Array Fabrication:A: The fabrication begins with a 500 pm-thick silicon wafer.B: Polyimide (PI) is spin-coated onto the silicon wafer, followed by baking and curing. This step is repeated to increase the thickness of the PI layer.C & D: After roughening the PI in a plasma asher, lift-off resist and positive photoresist are spin-coated onto the polyimide layer.E & F: Using either a hard mask and UV exposure or a UV (405 nm) direct-write laser, the desired electrode geometry is patterned onto the photoresist, and the photoresist is developed.G: After a sample descum was done in the plasma asher, the sample is metallized with titanium and gold using e-beam evaporation. Sputtering of titanium nitride (TiN) may be done too.H: Both photoresists and the excess metal are lifted off, leaving the desired electrode pattern on the polyimide thin film.4935-6609-1653v.lI: Another layer of polyimide is spin-coated to encapsulate the electrodes.J: Thick photoresist is spin-coated onto the polyimide encapsulation.K: Using either a hard mask and UV exposure or a UV (405 nm) direct-write laser, the desired geometry for the electrode openings is patterned onto the photoresist, and the photoresist is developed, creating a photoresist-based etch mask.L & M: The polyimide is dry etched using O2 and CF4 plasma etching, creating openings in the poly imide to expose the electrodes.N: The photoresist is removed from the sample.O: The polyimide thin film with the electrode array is released from the silicon wafer using an excimer laser;

[0041] Fig. 12 shows an embodiment of the Stentinel involving direct patterning of electrodes onto the stent (as opposed to being on a rolled film of polyimide). In this example, an expanded stent is shown with spine segments highlighted. 250 triplets of electrodes are shown on the stent. There is a spine containing segments onto which the ASIC can be bonded, while still enabling the stent structure to remain flexible. When the stent is crimped, the struts are crimped in a way that the spine segments are unperturbed.

[0042] Fig. 13 is a close up of electrode triplets on an exemplary stent structure;

[0043] Figs. 14A-14B show in A an angled view of an exemplary stent, and B shows an exemplary stent with a flexible ASIC bonded thereto;

[0044] Figs. 15A-15B show in A an exemplar}' flexible ASIC bonded to spine segments, and B shows a Stentinel mounted on a balloon catheter (expanded) for. e.g., deliver}’ to and expansion within blood vessel;

[0045] Fig. 16 shows an exemplary crimped stent.

[0046] Throughout the drawings, the same reference numerals, and characters, unless otherwise stated, are used to denote like features, elements, components, or portions of the illustrated embodiments. Moreover, while the present disclosure will now be described in detail with reference to the figures, it is done so in connection with the illustrative embodiments and is not limited by the particular embodiments illustrated in the figures and the appended paragraphs.DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS4935-6609-1653v.l

[0047] The following is intended to be a description of the exemplary embodiments of the present disclosure, and is not intended to limit the scope of the exemplary embodiments of the present disclosure.

[0048] An implantable active bioelectronics stent, comprising:a mechanically flexible package;an active integrated circuit (IC) chip associated with the flexible package; andone or more electrodes configured to sense biomarkers in a bloodstream and associated with the active IC chip, wherein the stent is powered by wireless powering and communicated with via electromagnetic or ultrasound energy.

[0049] In some embodiments, the one or more electrodes can effect electrochemical impedance spectroscopy of cells in the bloodstream.

[0050] In some embodiments, the one or more electrodes can effect electroporation of cells in the bloodstream.

[0051] In some embodiments, the powering of the device is through a near-field inductive coupling.

[0052] In some embodiments, the communication to and from the stent occurs over one or more impulse-radio, ultra-wide-band communications links.

[0053] In some embodiments, the stent is mounted on a bioresorbable scaffold.

[0054] In some embodiments, the stent is mounted on a non-bioresorbable scaffold.

[0055] In some embodiments, the bioresorbable scaffold is composed of poly -L -lactide.

[0056] In some embodiments, the scaffold contains open cells aligned with the arrangement of the electrodes.

[0057] In some embodiments, the bioresorbable scaffold contains dedicated attachment points for the flexible electronics package.

[0058] In some embodiments, the active IC chip is mechanically thinned so as to be mechanically flexible.

[0059] In some embodiments, the flexible package integrates therein or thereon a metallic coil.

[0060] In some embodiments, the active IC chip includes an antenna thereon or therein.

[0061] In some embodiments, an external wearable device is placed over a location of the stent.

[0062] In some embodiments, the electrodes are attached to the stent structure via a flexible polyimide package.4935-6609-1653v.l

[0063] In some embodiments, the electrodes are directly patterned onto said stent.

[0064] In some embodiments, the electrodes are arranged in triplets.

[0065] In some embodiments, the electrodes are arranged to effect electrochemical impedance spectroscopy detection of cells or biological components.

[0066] In some embodiments, the electrodes comprise an antifouling surface for proteins, optionally a zwitterionic sulfobetaine coating.

[0067] In some embodiments, the device is adapted to effect electroporation so as to reduce, prevent or reverse endothelialization when implanted in a blood vessel of a subject.

[0068] In some embodiments, the device comprises electrodes arranged to effect electroporation of cells travelling through the lumen of the stent.

[0069] In some embodiments, the device comprises at least a portion of arranged to effect electrochemical impedance spectroscopy (EIS) disposed on a proximal end or half of the stent and at least a portion of arranged to effect electroporation of cells travelling through the lumen of the stent disposed on a distal end or half of the stent.

[0070] In some embodiments, the device comprises an ASIC containing front-end analog circuits to support EIS detection and electroporation.

[0071] In some embodiments, the device is programmed to be capable of detecting CTC via EIS detection and electroporation of said CTC as it flows through the lumen of the stent.

[0072] In some embodiments, impedance of a cell travelling through a lumen of the stent is determined at 2 or more frequencies simultaneously and identified as a CTC when it meets preidentified impedance values for CTCs in the same species.

[0073] In some embodiments, impedance is determined using a three electrode arrangement.

[0074] In some embodiments, device is powered by, and / or communicated with, a wearable external relay station device.

[0075] In some embodiments, the electroporation is effected with pulsed current.

[0076] In some embodiments, the electroporation is effected with pulsed current with three phases.

[0077] In some embodiments, the electroporation is effected with pulsed current with an electroporation pulse durations of up to 375 ps.

[0078] A device comprising:an implantable active bioelectronics stent which comprises:a mechanically flexible package;4935-6609-1653v.lan active integrated circuit (IC) chip associated with the flexible package; andone or more electrodes configured to sense biomarkers in a bloodstream and associated with the active IC chip, wherein the device uses wireless powering and communication via electromagnetic or ultrasound energy’.

[0079] A kit comprising a device described herein, and an external wearable device for placing in close proximity with said device and communicating with said device. In some embodiments, the wearable device comprises a relay station.

[0080] A method of diagnosing metastatic potential in a subject comprising detecting a circulating tumor cell (CTC) in a bloodstream of a subject using the device described herein by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device, wherein presence of a CTC in the bloodstream is indicated by differential EIS trace indicative of a CTC and indicates metastatic potential, and wherein absence of detection of a CTC in the bloodstream does not indicate metastatic potential.

[0081] A method of reducing the likelihood of metastasis in a subject comprising detecting a circulating tumor cell (CTC) in a bloodstream of a subject using the device described herein by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device, wherein presence of a CTC in the bloodstream is indicated by differential EIS trace indicative of a CTC and indicates likelihood of metastasis, and wherein when the CTC is detected electroporating the CTC via the device.

[0082] A method of treating a hematological cancer in a subject comprising detecting a hematological cancer cell in a bloodstream of a subject using a device described herein by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though a stent of the device, wherein presence of a hematological cancer in the bloodstream is indicated by differential EIS indicative of a hematological cancer cell and, wherein when the hematological cancer cell is detected, electroporating said cell via the device.

[0083] A method of screening a candidate treatment for reducing or removing circulating tumor cells (CTCs) in a bloodstream of a subject using the device described herein by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device on a plurality of occasions during or subsequent to administration of the treatment, and quantifying if the number of CTCs detected in the bloodstream over the plurality of occasions is reduced during or subsequent to administration of the treatment, wherein presence of a CTC in the bloodstream is indicated by differential EIS trace indicative of a CTC.4935-6609-1653v.l

[0084] In some embodiments, the methods further comprise surgically implanting or delivering a stent of the invention into a blood vessel of the subject prior to detecting a CTC. In some embodiments, the stent implanted is an expandable stent, and the method optionally comprises expanding the stent once in situ. In some embodiments, the stent implanted is an expandable stent and is delivered via a ballon catheter. In some embodiments, the methods employ one stent with both EIS and electroporation capability. In some embodiments, the methods employ two stents, including at least one having at least EIS and the other having at least electroporation capability.

[0085] Fig. 1 shows a diagram of a non-limiting exemplary device, e.g., which can sometimes be termed as "Stentinel" herein (or, e.g., a '‘Stentinel system” including the relay station). The exemplary Stentinel can be deployed by conventional methods, for example, with a balloon catheter, and can include in some embodiments a polyimide package containing the ASIC of the exemplary Stentinel, electrodes, and wireless powering coil. In some embodiments, the exemplary Stentinel can be inserted into the cephalic vein in the upper arm with a balloon catheter. An active bioelectronic stent as disclosed herein can continuously scan the bloodstream for CTCs and liquid cancer cells, including CHCs, facilitating the entire blood system to be scanned, overcoming the traditional challenges associated with liquid biopsies. In some embodiments, the device / system can detect a predetermined biological marker via the EIS methodology described herein. In some embodiments, the predetermined biological marker is a cell type, ctDNA or exosomes. In some embodiments, the predetermined biological marker is a metabolite. In some embodiments, the predetermined biological marker is a blood lipid. In some embodiments, the CTC in the methods described herein is from a breast, prostate, colorectal, gastric (stomach), pancreatic, lung, or liver cancer. In some embodiments, the CTC in the methods described herein is EpCAM positive. In some embodiments, in the methods described herein, the Stentinel detects and or destroys CAFs (cancer-associated fibroblasts) and / or ( TAMs (tumor-associated macrophages). In some embodiments of the methods disclosed herein, the subject is a human.

[0086] In some embodiments, vessels are selected to be less than 5 mm in diameter and to be less than 2 cm from the surface of the skin. For example, thrombosis at such a chosen location should present very minimal risk to the patient. In some embodiments, vessels are selected to be less than 10, 9, 8, 7 or 6 mm in diameter. In some embodiments, vessels are4935-6609-1653v.lselected to be less than 1.9, 1.8., 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1 or 1.0 cm from the surface of the subject’s skin.

[0087] Any suitable location can be used. In non-limiting examples peripheral arteries, iliac vein, coronary’ artery, artery or vein of any specific organ. In some embodiments in treating a cancer or preventing or reducing metastasis the stent(s) can optionally be placed in a vascular vessel of the bloodstream of the organ or leaving the organ. In some embodiments the stent is placed in a blood vessel of a limb of the subject. In some embodiments the organ is a kidney, lung, liver, heart, brain, or pancreas.

[0088] In some embodiments, the subject is a human subject.

[0089] Stents can be any suitable size, especially for human subjects. In some embodiments, the stents are from 2.0 to 5.0 mm in diameter. In some embodiments, the stents are from 2 mm to 48 mm in length. In some embodiments, the stents are from 15 mm to 20 mm in length.

[0090] In some embodiments, more than one stent device is employed. For example, two stents in series can be employed. In some embodiments, the two stents communicate wirelessly with and / or are controlled by the same processor.

[0091] In some embodiments, the stent is not wired or physically connected to any device or component external to the body.

[0092] In some embodiments, the electrodes comprise any suitable biologically inert substances which are electrode material. For example, in some embodiments the electrodes can comprise titanium nitride. In some embodiments the electrodes can comprise platinum or nitinol.

[0093] The exemplary7systems and devices described herein, e.g. a Stentinel system, can be, or include, a wirelessly powered active device that is placed in the vascular system. Foer example, in some embodiments, in the cephalic vein in the upper arm with, e.g, entry at the antecubital fossa. In some embodiments, the system can be powered by an external battery-pow ered "relay station" which is worn over, or sufficiently close to, the site of the implant, and which, for example, also communicates with the device through a high-bandwidth wireless data link. The relay station is itself can be, in some embodiments, an 802.1 In WiFi device. This allows the stent to communicate wirelessly through this relay with software running on a processor, such as a smartphone, smartwatch, or desktop or laptop computer. In some embodiments, other WiFi devices are employed, e.g., 802.1 lac or 802.1 lax.4935-6609-1653v.l

[0094] Within the exemplary system, the scanning can be performed by electrodes positioned on the walls of the vein, or within the stent (e.g. on the lumen-side surface of the stent struts or splines), which perform electrochemical impedance spectroscopy (EIS) of cells or biological markers passing within, e.g.. approximately 50 mm of the vessel walls. EIS is known in the art, e.g. Magar et al, (2021), found on the world wide web at pmc.ncbi.nlm.nih.gov / articles / PMC8512860 / which is hereby incorporated by reference.

[0095] From the resulting current traces, e.g. performed at exemplary two frequencies such as, for example, 300 kHz and 10 MHz, distinctive different signatures of CTC / tumor cells (indicative of differences in size or impedance) can be identified, for example by machine learning (ML) or compared to predetermined characteristic / control EIS signatures that have been effected for specific biomarker types, CTC types etc. The detection of CHCs can be straightforward because of their anomalous size. In other embodiments, the CTCs, can also, or separately, be distinguished by their anomalous nuclear-to-cell ratios. Under software control, the same electrodes detecting the cell can be directed to irreversibly electroporate and kill or damage the detected cells. Alternatively, sets of electrodes of the stent which are “upstream"’ in the bloodstream orientation pf the stent once in place in the blood vessel can be employed to detect the CTCs (or cell of interest) and sets of electrodes of the stent which are "downstream" in the bloodstream orientation once in place in the blood vessel can be directed to irreversibly electroporate and kill or damage the detected CTCs / cells.

[0096] There has been extensive work looking at EIS for identifying CTCs in external microfluidic systems as a label-free detection approach. EIS generally relies on the differences in size and dielectric properties to distinguish CTCs from normal cells [see, e.g., Ref. 39], There have been several papers utilizing CMOS biosensors for EIS [see, e.g., Ref.40], which essentially duplicates hardware developed by previously [see, e.g., Refs. 41-43], In operation of the exemplary device according to the exemplary’ embodiment of the present disclosure, the use of EIS to distinguishing CTCs in impedance flow' cytometry can be employed set up wdth highly restricted channels. Refs. 44-50 are useful background work incorporated by reference herein. One of the important differences of the exemplary device according to the exemplary embodiments of the present disclosure from these flow cytometry systems can be that restrictive channels are not present with a vascular deployment. This can mean that the blood supply cycle passes the exemplary stent or Stentinel system a number of times (e.g., many times) to be fully analyzed. In some4935-6609-1653v.lembodiments, EIS can be effected by applying current through the '‘outer” two of a three electrode array and the current measured by the “inner” of the three electrode array. For example, see Fig. 3.

[0097] In addition to the EIS detection, the exemplary Stentinel system can electroporate cells. An ablation technique has been considered for solid tumors [see, e.g., Refs. 51-53], electroporation conditions that introduce cell death in CTCs have also been considered for various cell lines. What is important for an electroporation can be the use of pulses to overcome Debye length screening through the application of fields that are introduced so rapidly that they cannot be screened by counterions. Fields on the order of e.g., approximately 375-437.5 V / cm have been shown to produce irreversible electroporation (IRE) of CTCs while allowing normal cells to eventually recover [see, e.g., Refs. 54-56], In some embodiments fields of 350-450 V / cm are employed. Cell death by IRE usually occurs through apoptosis or necrosis induced by permanent membrane disruption, leading to irreparable loss of homeostasis [see, e.g., Ref. 57],

[0098] The exemplary system can include a custom flexible polyimide package (see Figs. 2a and 2b, for example) containing, e.g., the electrodes, wireless powering coil, and custom complementary metal-oxide-semiconductor (CMOS) application-specific integrated circuit (ASIC). The CMOS ASIC, attached to the package through, for example, gold-gold hybrid bonding, can itself be thinned to less than, e.g., about 10, 15, 20, 25 or 30 um such as to be mechanically flexible. In some embodiments, it is thinned to about 20 um. The ASIC can also contain the antenna for the custom impulse-radio ultra-wide-band transceiver that establishes the communications link between the stent and the relay station. This exemplary polyimide package can be attached to bioresorbable poly-L-lactide (PLLA) stent with openings aligned with the electrodes and dedicated attachment points for the polyimide, and can be expanded by a balloon catheter which holds the polyimide package patent with the vessel walls while it is endothelialized. In some embodiments, the stent is nonbioresorbable.

[0099] In some embodiments, the stent herein is a permanent (e.g., non-bioresorbable polymer). In some embodiments, the stent herein comprises electrodes disposed on the inner lumen-side surfaces of the stent. In some embodiments, the electrodes are disposed in a three-electrode arrangement or triplet of electrodes. In some embodiments, the stent is wireless and is wirelessly powered. In some embodiments the stent structure itself is non-metal (although the electrodes thereon can be metal). In some embodiments, an CMOS4935-6609-1653v.lASIC is integrated into the system on the stent. In some embodiments, the ASIC is directed to EIS detection of a predefined cell type or biomarker. In some embodiments, the ASIC is directed to electroporation of a predefined cell type. In some embodiments, the cell is a CTC.

[0100] In some embodiments, the flexible polyimide package is patterned and mounted directly on the stent structure itself, preferably following the struts of the stent, resulting in wiring and electrodes contained directly on the stent. In some embodiments of this design, the stent will have an ‘“open cell” design as shown in Fig. 9 / 10, which has the advantage of less length shrinkage when expanded from the crimped state. This design also allows the attachment points for a Stentinel ASIC to maintain their spacing between the crimped and expanded states.

[0101] On top of the stent struts, gold or other metal (in some embodiments, or alloy) interconnect (Fig. 10B). encapsulated in, e.g.. polyimide passivation, thus allowing the stimulation electrodes and wireless powering coil to be wired out from the Stentinel ASIC, which can be attached to the stent structure by the same approaches used for attaching the ASIC to the polyimide package in the first embodiment. Since the stent often will have a metal core, breaks in some embodiments are positioned in this metal structure to avoid the formation of eddy currents from the wireless powering signal.Exemplary7Electronics Design of Exemplary Stentinel System: ASIC Design

[0102] What can be important (but not necessarily essential) to an exemplary design of the exemplary Stentinel system can be that the ASIC includes the front-end electronics for both EIS and electroporation, data conversion, the wireless transceiver, the wireless powering circuit, and a digital controller. The exemplary chip itself can be designed in a TSMC 0.13 pm CMOS BCD process, leverage a significant part of the circuit infrastructure that has been developed for the BISC BCI device [see, e.g., Ref. 58], exemplary! component(s) are shown in Fig. 4 (along with others). For example, as shown in Fig. 4, an ASIC of the exemplary' stent system can contain all of the front-end analog circuits to support EIS detection and electroporation, as well as wireless powering circuits and an IR-UWB transceiver. The exemplary system can be powered by. and communicated with via. a wearable external relay7station device.

[0103] Analog front end (AFE). For example, the impedance of a cancer cell / CTC in the vessel can be measured at multiple (e.g., 2) frequencies simultaneously through an injection4935-6609-1653v.lof fully differential superimposed voltage carriers at, e.g., about 300 kHz and about 10 MHz on the outer electrodes (El and E3) in the half-bridge, three-electrode configuration as shown in Fig. 3, which shows an illustration of a differential EIS measurement of cells in a half-bridge topology from the three-electrode detection module, according to the exemplary embodiment of the present disclosure. Red blood cells can be treated as a "noise" background. Fig. 3 shows that simulation voltages are applied between Electrodes 1 (El) and 3 (E3) with the current measured through Electrode 2 (E2).

[0104] For example, with no cell causing an impedance imbalance at the measurement site, the fully differential injected voltage on electrodes El and E3 cancel each other out at electrode E2. When a cell passes over the measurement site an AC potential proportional to the impedance imbalance between electrodes E1-E2 and E2-E3 develops on the middle electrode. The resulting in-phase and quadrature components of the measured current on E2 constitutes the measured signal.

[0105] The injected voltage carriers are generated with a single fully differential 10-bit capacitive digital-to-analog converter (DAC). The spacing of the three-electrode sensing modules (see Figs. 2A and 2B) can be selected such that all can simultaneously inject the same frequencies with minimal interference. On average, in the blood vessel, sensed cells can complete a full traversal of the three electrodes, e.g., in about 2 msec. A sample rate of 10 kS / s, for example, can be allocated to each EIS measurement channel to capture both the in-phase (I) and quadrature (Q) components of the signal. A sample rate of 5-15 kS / s, for example, can be allocated to each EIS.

[0106] There are multiple exemplary approaches to the analog frontend, for example: (1) amplify both carriers with a wideband amplifier first, or (2) mix down the modulated carriers first. Due to the power constraints, using the mixer first approach can be preferable. While this can mean that each measurement site should use a number (e.g., four) transimpedance amplifiers (TIA) - one for the in-phase measurement and another for the quadrature measurement for both injection frequencies - instead of only one TIA as used in the amplifier-first topology, the reduced bandwidth requirements can result in a significant power savings. Each TIA can be followed by low-pass filter, e g., at about 2 kHz before being multiplexed into a single analog-to-digital converter (ADC). In other embodiments, a low pass filter of 1.5kHz to 2.5kHz is used. In other embodiments, any suitable low- pass filter is employed.4935-6609-1653v.l

[0107] The electroporation can be accomplished with, for example, pulses with three phases. The first two phases can be anodic or cathodic, generating currents from two on-chip regulated current sources. During these phases, switches that connect the stimulation electrode to the corresponding current source can be activated, allowing the cunent to flow from the source to the electrodes. Both anodic and cathodic current have the same amplitude which can be programmable from, e.g., about 12.5 pA to about 3.28 mA. In some embodiments, the anodic and cathodic current are programmable either independently or the same to from about 10 pA to about 3.75 mA. In some embodiments, the third phase can be for charge balancing, in which the electrodes used for stimulation can be grounded to ensure no accumulated charge on the electrodes. The duration of all three phases can be independently programmed, e.g., from 0 to 375 ps. In some embodiments, the duration of all three phases is independently programmed, e.g.. from 0 to 500 ps. The compliance voltages of the anodic and cathodic current sources can be but not limited to, e.g.. ±5 V. In some embodiments, the compliance voltages of the anodic and cathodic current sources are ±7.5 V.

[0108] Transit Time Calculations:

[0109] We assume a flow velocity in the cephalic vein to be 8.6 cm / s. and a parabolic, laminar flow profile where cells travelling at center of the vessel will travel more quickly than ones at the edge. A cell travels over Electrode 1 (El) to Electrode 2 (E2) (see Fig. 3) where cell detection is performed (e.g., distance of 60 pm), and then to Electrode 3 (E3) where electroporation is performed (e.g.. distance of 40 pm).

[0110] For X = 0 pm (touching electrodes):

[0111] - The time to travel over El and E2 for detection is 23.3 msec.

[0112] The time to travel from E2 to over E3 for electroporation is 15.6 msec.

[0113] - Total time to travel all 3 electrodes is 38.9 msec.

[0114] For X = 30 pm:

[0115] The time to travel over El and E2 for detection is 11.7 msec.

[0116] The time to travel from E2 to over E3 for electroporation is 7.8 msec.

[0117] - Total time to travel all 3 electrodes is 19.5 msec.

[0118] Comparison With Electronics: The electrodes can measure at a sample rate of 10 kS / s, corresponding to 0.1 ps per sample. The electroporation pulse durations can range between 0 and 375 ps. Thus, the identification and electroporation steps take time on the order of a microsecond while the cell transit times over the electrodes are in the order of4935-6609-1653v.lmilliseconds, i.e., Upon encountering a current trace with a pre-determined set of characteristics identifying a cancerous cell (or CTC), the stent can electroporate the targeted cells.

[0119] Exemplary Data Conversion. For example, in one exemplary embodiment, since a single 10 bit, lOMS / s ADC can be used to digitize all channels for transmission to the relay station, each of the 256 EIS channel can be allotted 40kS / s - lOkS / s for each of the in-phase / quadrature measurements at each frequency. Each of these signals can be oversampled at five times the bandwidth.

[0120] Exemplary Wireless Transceiver. According to an exemplary embodiment of the present disclosure, the exemplary wireless transceiver can include, e.g., of UWB antenna and transceiver circuits and can utilize, e.g., the BISC BCI design [see, e.g., Refs. 58 and 59], On-off keying (OOK) can be used for data modulation. The wireless transceiver can be designed for low-power, single-user, and short-range communication. The UWB antenna, having a differential-fed slot design [see, e g., Ref. 60] can be located on the Stentinel ASIC. To achieve this, exemplary multi-layer human head models can be provided that can incorporate various tissue characteristics, including tissue thickness [see, e.g., Ref. 61] and dielectric constants [see, e.g., Ref. 62], It is possible to perform electromagnetic simulations to determine the parameters of the UWB antenna embedded in a blood vessel. Given that both the UWB antenna and the transceiver circuits can be provided for the implant, they can be co-designed for optimal performance. For example, a 700-MHz bandwidth can be achieved from, e.g., about 3.6 to 4.3 GHz.

[0121] The UWB antenna can be shared by the transmitter and receiver circuits using two sets of NMOS switches. To minimize insertion loss at the working frequencies (e.g., about 3.6 to 4.3 GHz), a floating-body technique [see, e.g., Ref. 63] can be used which can take advantage of the deep p-well (DPW) in the CMOS technology. Resistors can be used in-series with their gates to reduce RF leakage to the DC control lines.

[0122] Duty-cycling is a proven technique to reduce power consumption of impulse radio UWB (IR-UWB) transceivers [see, e.g., Refs. 64 and 65], The exemplary transmitter can employ this technique by generating short RF pulses using a duty cycled LC complementary oscillator. The LC oscillator can be tuned to resonate with the UWB antenna at, e.g., about 4 GHz without a local frequency reference. A digital pulse generator can generate a number (e.g., two) short baseband pulses with 1.3 and 0.3 ns pulse width from an enable signal. When transmitting a data "1", e.g., the LC oscillator's tail current can4935-6609-1653v.lbe turned on, e.g., for 1.3 ns. The LC oscillator can deliver, e.g., about 1 V peak-to-peak differential signal into the UWB antenna when it is turned on. The exemplary transmitter can transmit, e.g., at maximum 108.48 x 106pulses per second and consume, e.g., about 39 pj for each pulse transmitted.

[0123] The exemplary receiver can have a non-coherent energy detector architecture for its low implementation complexity [see, e.g., Refs. 66 and 67], For example, with a noise floor (No) at body temperature of -173.6 dBm, a minimal signal-to-noise ratio (SNRREF) of 17 dB for a bit-error rate (BER) less than 1 x 1O'10

[0068] , path loss (PL) of about 50 dB (which is conservative), receiver noise figure (NF) of about 5 dB and data rate (DR) of 54.24 Mbps, the minimal transmit power (PTX) of the relay station can be given by:PTX - SNRREF + 101 og(DR) + PL + NF + No = -24.3 dBm.According to the FCC regulations on the UWB band [see, e g., Ref. 69], e.g., a maximum equivalent isotropic radiated power (EIRP) of about -41.3 dBm / MHz can be transmitted in free air and measured over a 1ms time window. For example, the maximum power (PTX MAX) allowed to transmit for, e.g., a 500-MHz bandwidth is given by:P X MAX=EIRP + BW = -14.3 dBm.

[0124] It is also possible to comply with the average power limit as PTX is less than PTX_MAX. In addition, e.g., the peak power limit of 0 dBm can be satisfied as the high-data-rate regime is operated [see, e.g., Ref. 67], Since there can be a -14.3 + 21.3 = 10 dB link margin, this exemplary design can be robust against manufacturing variations and variations in the distance between the implant and relay station.

[0125] The UWB receiver features an RF architecture similar to the architecture previously described [see, e g., Ref. 70], The exemplary signal processing can start with amplifying the input signal from the UWB antenna by a low-noise amplifier (LNA), which can comprise an input stage followed by, e.g., two or more RF amplifier stages that provide, e.g., about 57 dB of voltage gain with in-band noise figure less than, e.g., about 5 dB. A self-mixing mixer based on double-balanced Gilbert-cell can detect the signal's envelope, which can be low-pass filtered by a baseband amplifier. Further, e.g., a one-bit digitizer (1-b ADC) comprised of a sampling and subtraction stage followed by a StrongArm comparator [see, e.g.. Ref. 71] can determine the received bit (e.g., "0" or "1") based on a predefined voltage threshold.

[0126] In an exemplary embodiment of the present disclosure, a phase-locked loop (PLL) [see, e.g., Ref. 72] can be used to generate, e.g., a 208.96 MHz clock from, e.g., the4935-6609-1653v.l13.56 MHz powering frequency, which can be divided down to generate all the clocks used in the implant. A four-stage differential ring oscillator can used which facilitates a timing control at the resolution of, e.g., about 0.567 ns. An exemplary digital controller inside the wireless transceiver can control the start-up of the PLL and timing generation. The LNA can be powered on for, e.g., about 4.61 ns, and the mixer and based-band low-pass amplifier can be powered on for 6 ns for ever}718.44 ns. For example, the entire transceiver can consume, e.g., about 12.2 mW when transmitting, e.g., at 50% of "T's.

[0127] Exemplary7controller of Exemplary7Stentinel System. The exemplary digital controller on the exemplary Stentinel system is responsible for managing the wireless communication, device configuration, and dynamic data acquisition in a manner similar to the BISC BCI device [see, e.g., Refs. 58 and 59], The exemplary7digital controller can operate from, e.g., about a 1.5-V power domain on the chip and consumes a peak power of, e.g., about 10 mW during EIS recording. The exemplary controller can include a main decoder that can translate the instructions received by the wireless transceiver into actions on the device. The decoder can decode, e.g., only one instruction at a time and while an instruction is being executed, other instructions cannot be accepted by the controller.

[0128] The exemplary digital controller can interface to two or more specialized control blocks:Exemplary AFE control. The exemplary AFE control can set the static configuration for EIS recording and electroporation stimulation functions and outputs dynamic control signals that can control the switches in the AFE.Exemplary Communications control. This exemplary block can receive data samples from the AFE and packetize them for transmission via the wireless transceiver. During a recording, every7new sample of data can be immediately added to the next packet. When a packet is full, it can be passed to be streamed through the wireless transceiver, while the preparation of the next packet begins. The communication control can also be configured to packetize and transmit other types of data regarding general information on the current state of the device. The packetizing can follow7a specialized communication protocol provided for the exemplary7Stentinel system according to the exemplary7embodiment of the present disclosure.

[0129] The exemplary instruction set architecture (ISA) of the exemplary7controller can include a number of (e.g., seven) instructions that can be classified into three types: dynamic instructions, static instructions, and the query7configuration instruction. The4935-6609-1653v.lexemplary dynamic instructions can include recording, stimulation, power-on, and halt. Each of these exemplary instructions can initiate an event that can start and / or stop an ongoing process. The exemplary recording instruction can start a process that can updates the control lines going to the AFE which facilitates an exemplary’ EIS recording from, e.g., most or all 256 EIS modules on the exemplary' Stentinel system. It is also possible to record only from a designated subset of EIS modules at a higher sample rate. The exemplary stimulation instruction can initiate an electroporation between the configured electrodes of a specified three-electrode module. The exemplary power-on instruction can power on the ElS-level recording amplifiers, which should be done prior to the recording. The exemplary halt instruction can stop an ongoing recording.

[0130] The exemplar}’ static instructions can include, e.g., configuration and programming. The exemplary configuration instruction can program, e.g.. up to 64 bits of configuration in a given instruction. This exemplary configuration can include register(s) (which can be in total, e g., 122 bits), control the functioning of finite state machines (FSMs) in the controller and / or are distributed to other functional blocks on the Stentinel implant, such as, e.g., the WPT, the AFE, and the UWB transceiver in order to set longer-term states that are infrequently updated. The exemplary programming instruction can set the address of electrodes used for stimulation and their initial polarity7, along with additional configuration information that are stored in the registers of the exemplary controller. When the stimulation is initiated, this information can be used to gate specific control signals sent to the AFE. The exemplary query configuration instruction can facilitate a request to access to the contents of the configuration registers.

[0131] Exemplary Wireless poyver (WPT) circuits. The exemplar}’ WPT circuit can receive poyver from the relay station using, e.g., about 13.56 MHz inductive coupling, thereby converting the received power to, e.g., regulated ±5V and +1.5V supplies. RF inductive power received by the power coil can be first converted to a DC voltage using an active rectifier. This DC voltage, which can be anyyvhere between, e.g., about 2 and 3.3V to support full functionality of the exemplar}7Stentinel system, can be further converted to, e.g., about +1.5V, +5V and -5V supplies. To generate -5V, e.g., the exemplary rectifier output voltage can go through an additional stage prior to the regulation stage.

[0132] The output voltage level of the rectifier can be a function of the amount of power received by the implant of the exemplar}7Stentinel system and the amount of power consumed thereby. According to certain exemplary embodiments of the present disclosure,4935-6609-1653v.lboth factors can be dynamically changing. For example, the former can depend on the coil-to-coil alignment between the exemplary Stentinel system and the relay station headstage, and the latter can depend on the mode of operation of the exemplary' Stentinel system. For thermal and specific absorption rate (SAR) characterization, the exemplary Stentinel system can be in a recording state, which is likely the most power-intensive mode of operation.

[0133] According to certain exemplary' embodiments of the present disclosure, the power delivered to the device can be sufficient and no more than what is necessary' to support full operation, since the excess power received can it to be converted to heat. This can mean that under a certain preferred operation, the rectifier output voltage can be stabilized to near 2V. To achieve this, it is possible to periodically’ read out the rectifier output in a three-bit digitized format (for example, 0: 2V, 1: 2.1V, ..., 7: 2.7V) using the query configuration instruction to the implant of the exemplary Stentinel system and adjust the magnitude of transmitted power from the relay station.

[0134] Exemplary coil-to-coil link efficiency can be an important metric in an inductive WPT design. A further efficient link can facilitate a longer operating distance, as well as maintain the power radiated by the headstage to stay below the SAR safety exposure limit (e.g., 1.6 W / kg) set by the FCC [see, e.g.. Ref. 73], With an assumption of an ideal conjugate-matched impedance on the transmitting side (e.g., Port 1) and, e.g., about a 60 ? load on the receiving side (e.g., Port 2) which can be the periodic steady-state (PSS) simulated linear-load equivalent of the time-vary ing rectifier input impedance, a simulation S21 and power transfer efficiency (PTE) can be provided, as shown in Fig. 5. The exemplary Stentinel system can include the coil on the implant that is curved (see Fig. 5), thereby reducing its effective area for flux linkage, which can be maximized by having, e.g., the coil unroll to exactly the circumference of the vessel.Exemplary Polyimide packaging manufacture and assembly

[0135] An exemplary device fabrication can include, e.g., the fabrication of the flexible poly imide package with integrated electrodes, the attachment of the integrated circuit to this package, and the attachment of the Abbott Esprit polymeric stent to the polyimide package.

[0136] Exemplary Fabrication of the polyimide package. According to an exemplary embodiment of the present disclosure, a silicon wafer can be used as a earner for the polymeric stent fabrication. A layer of PI2611 polyimide can be spuncast and cured on the silicon wafer. An 18nm / 200nm titanium / gold metal interconnect layer can then be patterned4935-6609-1653v.lfollowed by additional passivation layer of polyimide for a resulting total substrate thickness of 7 pm. This interconnect layer facilitate the pads of the ASIC to connect to the 768 electrodes, and the wireless powering coil can also be formed in this interconnect layer. There can be enough sufficient routing on a single layer of interconnect to provide the necessary connections. The passivation layer can be etched over the pads for connection to the ASIC. To ensure a robust electrical connection to the underlying metal layer, 1-pm-thick gold pillars can be deposited with sputtering or electroplating to provide via metal from the package to the pads of the ASIC. The package can then be flipped over for electrode fabrication. Vias through the substrate to the interconnect layer for the electrodes can be fabricated with a secondary layer of 8nm / 200nm titanium / gold, that can be deposited to form the foundation for the electrode layer, which can be either sputtered TiN or laminated SWCNT films. The overall dimension of the polyimide package for the 4-mm-diameter vessel can be, e.g., 1.5 cm x 6.6 mm. Also, an exemplary fabrication is shown in Fig. 1 1.

[0137] Exemplary Attachment of the ASIC. According to an exemplary embodiment of the present disclosure, the pads of the 1.5 cm x 1.5 mm ASIC are electrolessly plated with Ni-Au pads that extend over the height of the passivation. There are then two approaches that can be used for attachment of the ASIC, an anisotropic conducting film (ACF) with 5-pm conductive balls or Au- Au hybrid bonding. In the case of ACF, it can provide an adhesive underfill and conductive interface to hold the chip in place. For the latter, it is possible to utilize thermocompression and / or thermosonic bonding at temperatures of approximately about 100°C. In both exemplary cases, the Finetech Fineplacer Lambda tool can be used to flip-chip position the ASIC over the pads of the package and perform the bonding After the bonding, the chip can be thinned to approximately, e.g., 20 pm using, e.g., an Allied Hi-Tech milling tool and / or an additional layer of PI2611 polyimide can be spuncast over the ASIC for encapsulation.

[0138] It can be noted that polyimide has received regulatory approval for < 30-day implantation by NeuroOne for a brain implantation application. Longer term implantation approval is forthcoming likely imminently which should provide this material an good choice for the use in the exemplary Stentinel system.

[0139] Attachment of the bioresorbable stent. In one example, the polyimide package can be attached to the PLLA stent linearly at the dedicated attachment points on the stent, e.g., along the ASICs 1.5-cm linear extent with a biocompatible epoxy.4935-6609-1653v.lStent design

[0140] An exemplary stent design, shown in Fig. 9, has two significant modifications to best function as a support scaffold for the Stentinel electronics package: 1) a set of large open cells (openings) aligned with the arrangement of the electrodes, and 2) a set of dedicated attachment points for the polyimide roll. A conventional stent design results in a significant number of electrodes being blocked, which interferes with EIS measurements. The proximity of the stent struts to the electrodes results in flow disturbances that can affect the EIS measurements too. To address this, an exemplary' custom stent design incorporates large open cells that are aligned with the arrangement of the electrodes to drastically lower the number of electrodes being blocked, as well as create a larger distance on average between the electrodes and the stent struts to lower the chance of flow disturbances affecting the EIS measurements. Non-bioresorbable stent materials include PBMA and PVDF-HFP or durable fluoropolymer(s). Bioresorbable stent materials include PLLA. In either case, it is preferable that the polymer material is biocompatible. In some cases, the stent may have a metal or alloy base structure which is coated with a polymer (e.g., cobaltchromium). In some cases, the stent does not have a metal or alloy base structure. In some cases, the stent comprises a durable polymer surface and / or structure.

[0141] In some embodiments, the stent comprises electrodes - e.g„ in triplets or as pairs. In some embodiments, the stent can comprise 256 electrodes.

[0142] Furthermore, an exemplary custom stent design can incorporate horizontally aligned attachment points for the polyimide or polyimide-comprising package. With a conventional stent design, the polyimide package would have to be bonded to the stent crowns (peaks) or stmts. This is not ideal given these are either regions of high stress or are areas which change in terms of angle and relative position during expansion, so bonding polyimide at these locations may compromise the mechanical integrity’ of the stent. The custom stent’s horizontally aligned attachment points enable the stent to expand with lowered mechanical interference from the polyimide package, and the larger attachment surface area can increase the bonding strength between the stent and polyimide package.Exemplary Electrode design and "defouling" procedures

[0143] Exemplary Micro-electrodes for sensing and electroporating cancer cells. Non-Faradaic electrodes can be used for the exemplary’ Stentinel system to reduce the effect of4935-6609-1653v.lelectrochemical reactions at the electrodes. Lower electrode impedance (e.g., higher capacitance) can boost signal levels in EIS recording and improve the effectiveness of stimulation for electroporation. In some embodiments of all the designs herein, both sputtered TiN electrodes and electrodes based on laminated mats of single-walled carbon nanotubes (SWCNT) are utilized. In some embodiments of all the designs herein, sputtered TiN electrodes are employed. In some embodiments of all the designs herein, SWCNT electrodes are employed. Both these electrode materials can provide a "roughened" surface which significantly increases effective surface area. The SWCNT electrodes can be based on the fabrication of free standing films of pristine SWCNTs (no binder / surfactant), formed by a liquid-air self-assembly process. The measured electrode impedance of, e g., about 20 pm x 20 pm TiN electrode is indicated in Fig. 6. In this case, the electrodes can have an impedance of about, e.g., 205 kQ at 1 kHz. which can correspond to a capacitance of, e.g., about 0.77 nF. Results on the SWCNT electrodes indicate that they can further reduce the impedance to, e.g., about 2.5 k at 1 kHz, corresponding to a capacitance of, e.g., about 64 nF. In other embodiments, any known suitable electrode material is used,

[0144] Exemplary Strategies for mitigating electrode biofouling and endothelialization of electrodes. Biofouling due to protein adsorption can dramatically increase electrode impedances over time. To address this issue, it is possible to functionalize the electrodes of the exemplary stent systems with anti-fouling reagents, For example, zwitterionic sulfobetaine coatings that have proven anti -fouling performance [see, e.g., Refs. 74 and 75], For the SWCNT electrode surface, sulfobetaine moieties can be chemically grafted [see, e.g., Ref. 76] to achieve a more robust electrode functionalization than the more commonly used physical adsorption methods. To accomplish this, sultone functionalized silane can be synthesized. Next, the electrodes can be electrochemically activated by, e.g., applying about +1.5V in 0.5M sodium hydroxide electrolyte relative to an Ag / AgCl reference to create hydroxyl groups on the SWCNT surface. The electrodes can be washed thoroughly and then exposed to the sultone functionalized silane for grafting the silane to the SWCNTs via condensation reaction.

[0145] Endothelialization of electrodes can be another concern. To address this issue, if desired, it is possible to rely on non-thermal electroporation to irreversibly rupture endothelial cells and other cells (e g. muscle cells) that tend to grow over the electrodes. Studies reveal that pulsed electrical fields irreversibly electroporate such cells and is a safe4935-6609-1653v.ltechnique causing no harm to the blood vessel and to the overall health of the subject [see, e.g., Ref. 77-80],Exemplary Relay station design

[0146] The relay station can both wirelessly power the implant and transfer data between the exemplary stent system and a computer base station, which can take the form of, e.g., a smartphone or a smart tablet. There can be a number of exemplary embodiments of the relay station, e.g., Generation-1 and Generation-2 versions. The exemplary Generation-1 (Gen-1) version can be a two-part system that includes a headstage and a processor module, which can be designed from commercial off-the-shelf components. The exemplary7Generation-2 (Gen-2) version can have only the headstage, and can rely on a custom ASIC design to further consolidate the volume required for the relay station. Fig. 7 shows exemplary generations of the relay station.

[0147] Exemplary Generation- 1 relay station design. In this exemplary embodiment, the headstage can have a wearable form factor with a printed circuit board (PCB) stack-up that can include a powering coil and ultra-wideband (UWB) antenna. For wireless powering, it is possible to use a tunable power amplifier to drive a custom spiral coil at, e.g., about 13.56 MHz that can inductively couple to the receiving coil on the implant. For the exemplary data telemetry, it is possible to use an impulse-radio ultra-wideband (IR-UWB) transceiver centered, e.g., at about 4 GHz with on-off-keying (OOK) modulation, where a custom dipole antenna on the headstage can communicate with a monopole antenna on the implant.

[0148] The exemplary processor module can control the headstage through a standard high-definition-multimedia-interface (HDMI) cable. The processor module can power and configure PCB components on the headstage, send queries and commands to the implant, and receive responses and recorded data from the implant. Recorded data can be saved in the processor module's non-volatile memory and / or re-directed to the computer base station over wired or wireless Ethernet. The exemplary processor module can be based on a Xilinx Zynq-7000 system-on-chip (SoC) that can include a processing system (PS) unit and a programmable logic (PL) unit. The PS integrates a dual-core ARM Cortex-A9 processor, can run the Linux operating system, and interface with a secure digital (SD) card used as the main non-volatile memory. The PL can integrate specialized hardware to handle the bitstream of recorded EIS data and store it in the ARM processor's main memory. In addition, the specialized hardware on the PL can embed a high-level application4935-6609-1653v.lprogramming interface (API) to generate and deliver pre-configured sequences of timesensitive commands to the implanted stent. The API can be reconfigurable, leveraging its implementation on the PL, thus facilitating it to be updated at any time.

[0149] The exemplary bi-directional communication protocol can, for example, use a custom 125-bit packet tailored to accommodate the sampling rate and ADC resolution of the exemplary stent system. The use of a custom protocol between the exemplary stent system and the relay station can provide a more energy -efficient operation of this link when compared to the use of standard protocols such as Bluetooth or 802.11. Moreover, radio transmission between the implant of an exemplar}' stent system and the relay station should be over a few centimeters, which allows the exemplary stent system to be a high-bit-rate radio transceiver while consuming less than 100 mW of total power (e.g., less than 13 mW peak power in the transceiver itself). The entire relay station can be battery-powered from, e.g., two Li-ion 18650 batteries, which can provide 5000 mAh of battery life. This facilitates the relay station to actively power one implant for about 24 hours, allowing for continuous EIS sensing across all 256 electrodes and infrequency electroporation events.

[0150] Exemplar}' Generation-2 relay station design. For exemplary Generation-2 relay station, it is possible to combine the processor board with the electronics on the head-stage board. The processor board can use, e.g., a Snickerdoodle board, which has considerably more function that is required by the current relay station. This can be replaced by a Zync SOC (e.g., FPGA and ARM processor) with a ATWILC3000 Wi-Fi module, allowing the software development to translate to the Generation-2 design. The compactness of the exemplary design can integrate most or all of the analog and mixed-signal functionality onto a custom ASIC. With a custom board and chip, the exemplary antenna board and processor board electronics can be combined onto a single rigid board, as shown in Fig. 7. The compact exemplary Generation-2 relay station design can make it wearable. The batter}' in this exemplary case can be, e.g.. eight Enovix EXTA-351830 batteries with a 3.5 mm x 18 mm x 30 mm form factor and a capacity of 250 mAh each.Exemplary Software and firmware design

[0151] Exemplary Core software architecture. Some of the core operations of the transceiver interface can be implemented in PL on the relay station. The software of an exemplary stent system, built on to top of this, can be based on an API to the instruction set of the stent controller as show n in Fig. 8. It is possible to implement a Python software4935-6609-1653v.lframework that can use this API and run under the Linux operating system using PYNQ to control the specialized hardware in the PL of the relay station. To support an interactive user environment, a graphical user interface (GUI) can run on the computer base station in the form of a smart phone or desktop computer and interact with the Python framework on the relay station via a RESTful API. The GUI can interact with the relay station through the high-level API. The software for an exemplary stent system can have a diagnostic mode in which most or all of the recorded EIS data from the exemplary' stent system is available in real-time and stored in the cloud-based storage, for example. A machine-learning inference engine analyzing the EIS data, as described herein, can be implemented first in software. It can also be moved to the PL of the relay station to facilitate the latency for detection to be reduced, enabling a real-time electroporation control.

[0152] Exemplary' Machine-learning procedures. To analyze the data generated from the EIS sensing as, it is possible to utilize the deep learning models to leam the vectorial representations of data and then make predictions of the cell size, nuclearcytoplasmic (N:C) ratio, and distance to electrodes. For each candidate cell, the input to the model can include the in-phase and quadrature components at three different frequencies. The initial data can be gathered from simulation with various cell sizes, N:C ratios, and distances. In some embodiments, a transformer-based model is used to leam the representation of the in-phase and quadrature component at each frequency. In some embodiments, extraction of heuristic features such as, e.g., the start value, peak value, and peak value coordinates, is employed. In some embodiments, the representations for each frequency and the heuristic features can be combined in a fully connected layer for final predictions. In some embodiments, other shallow machine learning methods are used, such as, e g., support vector machine (SVM) for comparison.

[0153] When the cell line data is available, it is possible to conduct a transfer learning to update the exemplary model and adapt to the new data. In addition, to enhance the model capacity and interpretability, in some embodiments, an interpretable "foundation model" is provided (e.g., called VQ-Shape) by pretraining on both simulation data and cell line data and extracting more shape patterns. This can provide better discrimination.

[0154] In some embodiments, the data generated from the EIS sensing is compared to predetermined data or models obtained previously and interpreted by machine learning. The predetermined data or models in some embodiments can be predetermined for two or more different frequencies, or three different frequencies, and optionally the in-phase and4935-6609-1653v.lquadrature components of the different frequencies used, so as to predict and desired identification parameter(s), e.g., cell size, nuclear: cytoplasmic (N:C) ratio, and distance to electrodes for cells such as e g., CTCs. The predictive ML model can be obtained beforehand for CTCs of the species in general (e.g., human) or for specific CTC type for a specific tumor (e.g., human lung, or breast, or prostate cancer, etc., or another cancer described herein), specific hematological cancer cells etc. as desired. In some embodiments, the methods employ such predetermined predictive models for data comparison or assessment. In some embodiments, the methods further comprise obtaining or having obtained such predetermined predictive models for data comparison or assessment. In some embodiments, a transformer-based model is or has been used to learn the representation of the in-phase and quadrature component at each frequency. In some embodiments, extraction of heuristic features such as, e.g., the start value, peak value, and peak value coordinates, is or has been employed. In some embodiments, the representations for each frequency and the heuristic features are combined in a fully connected layer for final predictions. In some embodiments, other shallow machine learning methods are used, such as, e.g., support vector machine (SVM) for comparison. The algorithm may employ any suitable neural network, as also described herein. CNNs or DNNs may be employed in some embodiments.

[0155] In some embodiments, the models are pretrained on actual cells from a human subject. In some embodiments, the models are pretrained on cells from a cell line. In some embodiments, the cells lines are human cell lines.

[0156] In some embodiments of the methods herein in regard to cancer, the cancer is a melanoma, a glioblastoma, a head and neck cancer or a lung cancer. In some embodiments, the cancer is a bladder cancer, kidney cancer, Hodgkin lymphoma, esophageal cancer, gynecologic cancers, pancreatic cancer, or a renal cell carcinoma. In some embodiments, the cancer is a cancer of a breast, nasophary nx, pharynx, lung, bone, brain, sialaden, stomach, esophagus, testes, ovary, uterus, endometrium, liver, small intestine, appendix, colon, rectum, bladder, gall bladder, pancreas, kidney, urinary bladder, breast, cervix, vagina, vulva, prostate, thyroid or skin, head or neck, or is a glioma.

[0157] In some embodiments, the cancer is a hematological cancer. In some embodiments, the cancer is an acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, a multiple myeloma, or a B-cell lymphoma.

[0158] In some embodiments, the models identify cells having a higher N:C ratio and / or larger size than is obtained for a population of non-malignant cells of the same cell type. In4935-6609-1653v.lsome embodiments, threshold values are identified forN / C ratio and / or cell size at or above which a cell is identified as malignant (cancerous).

[0159] In this description, numerous specific details have been set forth. It is to be understood, however, that implementations of the disclosed technology can be practiced without these specific details. In other instances, well-known methods, structures, and techniques have not been shown in detail in order not to obscure an understanding of this description. References to "some examples," "other examples," "one example," "an example," "various examples," "one embodiment," "an embodiment," "some embodiments," "example embodiment," "various embodiments," "one implementation," "an implementation," "example implementation," "various implementations," "some implementations," etc., indicate that the implementation(s) of the disclosed technology7so described may include a particular feature, structure, or characteristic, but not every implementation necessarily includes the particular feature, structure, or characteristic. Further, repeated use of the phrases "in one example," "in one exemplary embodiment," or "in one implementation" does not necessarily refer to the same example, exemplary embodiment, or implementation, although it may.

[0160] As used herein, unless otherwise specified the use of the ordinal adjectives "first," "second," "third," etc., to describe a common object, merely indicate that different instances of like objects are being referred to, and are not intended to imply that the objects so described must be in a given sequence, either temporally, spatially, in ranking, or in any other manner.

[0161] While certain implementations of the disclosed technology have been described in connection with what is presently considered to be the most practical and various implementations, it is to be understood that the disclosed technology' is not to be limited to the disclosed implementations, but on the contrary, is intended to cover various modifications and equivalent arrangements included within the scope of the appended numbered paragraphs. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

[0162] The foregoing merely illustrates the principles of the disclosure. Various modifications and alterations to the described embodiments will be apparent to those skilled in the art in view of the teachings herein. It will thus be appreciated that those skilled in the art will be able to devise numerous systems, arrangements, and procedures which, although not explicitly shown or described herein, embody the principles of the disclosure and can be4935-6609-1653v.lthus within the spirit and scope of the disclosure. Various different exemplary embodiments can be used together with one another, as well as interchangeably therewith, as should be understood by those having ordinary' skill in the art. In addition, certain terms used in the present disclosure, including the specification and drawings, can be used synonymously in certain instances, including, but not limited to, for example, data and information. It should be understood that, while these words, and / or other words that can be synonymous to one another, can be used synonymously herein, that there can be instances when such words can be intended to not be used synonymously. Further, to the extent that the prior art knowledge has not been explicitly incorporated by reference herein above, it is explicitly incorporated herein in its entirety. All publications referenced are incorporated herein by reference in their entireties.

[0163] Throughout the disclosure, the following terms take at least the meanings explicitly associated herein, unless the context clearly dictates otherwise. The term "or" is intended to mean an inclusive "or." Further, the terms "a," "an," and "the" are intended to mean one or more unless specified otherwise or clear from the context to be directed to a singular form.

[0164] This written description uses examples to disclose certain implementations of the disclosed technology, including the best mode, and also to enable any person skilled in the art to practice certain implementations of the disclosed technology, including making and using any devices or systems and performing any incorporated methods. The patentable scope of certain implementations of the disclosed technology is defined in the numbered paragraphs, and may include other examples that occur to those skilled in the art. Such other examples are intended to be within the scope of the numbered paragraphs if they have structural elements that do not differ from the literal language of the numbered paragraphs, or if they include equivalent structural elements with insubstantial differences from the literal language of the numbered paragraphs.

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Claims

1. Claims:

1. An implantable active bioelectronics stent device, comprising:a mechanically flexible package;an active integrated circuit (IC) chip associated with the flexible package; and one or more electrodes associated with the active IC chip configured to sense biomarkers and / or cells, and / or optionally electroporate cells, in a bloodstream, wherein the stent is optionally powered by wireless powering and / or communicated with via electromagnetic or ultrasound energy.

2. The device of Claim 1, wherein the one or more electrodes can effect electrochemical impedance spectroscopy of cells in a bloodstream passing through said stent.

3. The device of Claim 1 or 2, wherein the one or more electrodes can effect electroporation of cells in a bloodstream passing through said stent.

4. The device of Claim 1, 2 or 3, wherein the powering of the device is through a nearfield inductive coupling.

5. The device of any of Claims 1-4, wherein the communication to and from the stent occurs over one or more impulse-radio, ultra- wide-band communications links.

6. The device of any of Claims 1-5, wherein the stent comprises a non-bioresorbable scaffold.

7. The device of any of Claims 1-5, wherein the stent comprises a bioresorbable scaffold.

8. The device of Claim 7, wherein the bioresorbable scaffold is composed of poly-L-lactide.4935-6609-1653v.l9. The device of any of Claims 1-8, in which the scaffold contains open cells aligned with the arrangement of the electrodes.

10. The device of any of Claims 1-8, in which the bioresorbable scaffold contains dedicated attachment points for the mechanically flexible package.

11. The device of any of Claims 1-9, wherein the active IC chip is mechanically thinned so as to be mechanically flexible.

12. The device of any of Claims 1-10, wherein the flexible package integrates therein or thereon a metallic coil.

13. The device of any of Claims 1-11, wherein the active IC chip includes an antenna thereon or therein.

14. The device of any of Claims 1-12, wherein an external wearable device is placed over a location of the stent.

15. The device of any of Claims 1-14, wherein the electrodes are attached to the stent structure via a flexible polyimide package.

16. The device of any of Claims 1-14, wherein the electrodes are directly patterned onto said stent.

17. The device of any of Claims 1-16, wherein the electrodes are arranged in triplets.

18. The device of Claim 17, wherein the electrodes are arranged to effect electrochemical impedance spectroscopy detection of cells or biological components.

19. The device of any of Claims 1-18, wherein the electrodes comprise an antifouling surface for proteins, optionally a zwitterionic sulfobetaine coating.4935-6609-1653v.l20. The device of any of Claims 1-19, wherein the device is adapted to effect electroporation so as to reduce, prevent or reverse endothelialization when implanted in a blood vessel of a subject.

21. The device of any of Claims 1-20, comprising electrodes arranged to effect electroporation of cells travelling through the lumen of the stent.

22. The device of Claim 21. comprising at least a portion of arranged to effect electrochemical impedance spectroscopy (EIS) disposed on a proximal end or half of the stent and at least a portion of arranged to effect electroporation of cells travelling through the lumen of the stent disposed on a distal end or half of the stent.

23. The device of Claim 22, wherein the device comprises a CMOS ASIC containing front-end analog circuits to support EIS detection and electroporation.

24. The device of any of Claims 18-23, wherein the device is associated with programming capable of detecting CTC via EIS detection and electroporation of said CTC as it flows through the lumen of the stent.

25. The device of Claim 24, wherein impedance of a cell travelling through a lumen of the stent is determined at 2 or more frequencies simultaneously and identified as a CTC when it meets preidentified impedance values for CTCs in the same species.

26. The device of Claim 24, wherein impedance is determined using a three electrode arrangement.

27. The device of any of Claims 1-26, powered by, and / or communicated with, a wearable external relay station device.

28. The device of any of Claims 20-27, wherein the electroporation is effected with pulsed current.

29. The device of any of Claims 20-28, wherein the electroporation is effected with pulsed current with three phases.4935-6609-1653v.l30. The device of any of Claims 20-27, wherein the electroporation is effected with pulsed current with an electroporation pulse durations of up to 375 ps.

31. A device comprising:an implantable active bioelectronics stent which comprises:a mechanically flexible package;an active integrated circuit (IC) chip associated with the flexible package; andone or more electrodes configured to sense biomarkers in a bloodstream and associated with the active IC chip, wherein the device uses wireless powering and communication via electromagnetic or ultrasound energy.

32. A kit comprising the device of Claim 1 or 31, and an external wearable device for placing in close proximity with said device and communicating with said device.

33. The kit of Claim 32, wherein the wearable device comprises a relay station, optionally a wireless relay station which can power the active bioelectronics stent.

34. A method of diagnosing metastatic potential in a subject comprising detecting a circulating tumor cell (CTC) in a bloodstream of a subject using the device of any of Claims 1-31 by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device, wherein presence of a CTC in the bloodstream is indicated by differential EIS indicative of a CTC and indicates metastatic potential, and wherein absence of detection of a CTC in the bloodstream does not indicate metastatic potential.

35. A method of reducing the likelihood of metastasis in a subject comprising detecting a circulating tumor cell (CTC) in a bloodstream of a subject using the device of any of Claims 1-31 by performing electrochemical impedance spectroscopy on blood of the bloodstream passing though the device, wherein presence of a CTC in the bloodstream is indicated by differential EIS indicative of a CTC and indicates likelihood of metastasis, and wherein when the CTC is detected electroporating the CTC via the device.4935-6609-1653v.l36. The method of Claim 34 or 35 further comprising surgically implanting the stent prior to detecting a CTC.

37. The method of Claim 36, wherein the stent implanted is an expandable stent, and optionally comprising expanding the stent once in situ.4935-6609-1653v.l

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