(19) World Intellectual Property Organization International Bureau = (43) International Publication Date 09 November 2023 (09.11.2023) ‘TERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) TTA WA (40) aternational Publication Number WO 2023/214156 Al WIPO|PCT (61) International Patent Classification: G16B 5/30(2019.01) GUI 33400 2006.01) AGIBS100;200691) GSB 4020 201901) (21) International Application Number: PCTIGB2N20SI15S 2) International Fling Date: (02 May 2023 (02 05.2023) (25) Filing Language: English (26) Publication Language: English (80) Priovty Dat 22068364 (03 May 2022 (03052022) GB (71) Applicant: THE UNIVERSITY OF BATH [GBIGB}; Re- search Commercialisation Manager, Research & Innovation Services, Wessex House 520, Claverton Dowa, Bath and North Eat Somerset BAZ TAY (GB) (72) Inventors: NOGARET, Alain; The University of Bath Research & Innovation Services, Wessex. House 520, CCaveron Down, Bath and North East Somerset BA2 TAY (GB). WELLS, Stephen; The University of Bath, Re- search & Innovation Services, Wessex House 5.20, Claver- ton Down, Bath and North Eas Somerset BA2 TAY (GB), ‘TAYLOR, Joseph; The Unversity of Bath, Research ie novation Services, Wessex House $20, Claveron Down, Bath and Nosh East Somerset BA2 TAY (GB). MOR- RIS, Paul: Pigsiology Building, Depactment of Plysiol- ogy. Development and Neuroscience. University of Cam- be, Downing Place, Cambridge CB2 3EG (GB) (74) Agent: HUDSON, Daniel: Potter Clarkson, Chapel Qua (er, Mount Street, Nottingham NGI 6H1Q (GB), fy Recewe menbrane votage 10098, Vous.) ‘Sel membrare vologe variable = membrane voltage reading at every Mime points (69) Title: DETERMINING BIOLOGICAL PARAMETERS OF A CELL Seti ostinato of a2 | oy Veet: X 170.8) Fig. 1 Output a (57) abstract: The disclosure relates toa compute-inplemented meld for determining values of biological parameters ofa cel, $B comprising tecevng meminne volag eatingso he ox conesponding oN tine pols fora steve compring ie bola °G pametr anda pray ofinrdopende and tine-dpene ste vals, incing 2 nembrnevoling varie sting an “SF initial guess of the state vector and parameters; performing an optimization of the state vector by: setting the membrane voltage variable Ty tobe equa wa coresponding membrane vole reading eta mite of ine pins iin he Nine ps, determining a ped S saluc ofthe state veetorby opimizingan objective function that operates on values of te membrane voltage viable and corresponding, Inbranevalag eadigs, an ean the optinzaton wi he nial eat othe sae vets to he wpe vale of these vestor and he membrane vg varie sto eel a coresponngmenbrane vakage reading tee ine pois 23, Ss a ° S 1Comtinued on nex page}WO 2023/214156 A1 (MMIII (81) Designated States (unless otherwise indicated for every Jind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CV, CZ, DE, DI, DK, DM, DO, DZ, EC, EE, EG. ES, F, GB, GD, GE, GH, GM, GT, HN, HR. HU, D-IL. IN, 1Q, IR IS. T.JM, JO. JP KE, KG, KL KN, KP. KR, KW, KZ, LA, LC, LK,LR,LS, LU, LY, MA, MD, MG, MK, MN, MU, MW, MX, MY, MZ, NA, NG. NI, NO, NZ, OM, PA, PE, PG, PHL, PL, PT, QA. RO, RS, RU. RW. SA, SC, SD, SE. SG, SK, SL, ST SV, SY, TH, TLIM.TN, TR. TT. TZ, UA, UG, US, UZ, VC, VN, WS, ZA2M,2W. ey Designated States (unless otherwise indicated, for every Jind of regal protection avaiable: ARIPO (BW, CY, GH, GM, KE, LR, LS, MW, MZ,NA, RW, SC, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TH, TM), European (AL, AT, BE, BG, Cit, CY, CZ. DE, DK, EE, ES, Fl, FR, GB, GR. HR, HU, IE, 1S,IT, LT, LU, LV, MC, ME, MK, MT, NL, NO, PL, PE, RO, RS, SE, SISK, SM, TR), OAPI BF. BI, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, T) Published: = with inernational search report (Art 21(3) ‘win the Nie poisWO 2023/214156 PCTIGB2023/051155 DETERMINING BIOLOGICAL PARAMETERS OF A CELL Field The present disclosure relates to 2 computer-implemented method for determining values of biological parameters of a cell, and in particular to biological parameters related to one or more ion channels of a neuron. Background Channelopathies in which certain ion channels are over-expressed or absent occur in many neurological diseases and particularly in brain cancer and Alzheimer disease (AD). Hidden mutations in ion channels at the onset of differentiation pathways result in anomalous electrical activity which is sometimes accessible to measurement but is often too subtle to be detected and diagnosed. In addition, few of the mutations that may be revealed by genetic sequencing (transcriptomics, patch-sequencing) affect electrical function. Which genes affect electrical activity, and how, is largely unknown. Therefore, there is a need for a method capable of identifying the mutations responsible for the changes in electrical activity to develop targeted therapies and stem the decay of cognitive faculties in Alzheimer disease or brain cancer, for example, Several hypotheses exist as to which gene might be relevant to the growth and proliferation of gliomas in brain cancer. Tt is known that over-expression of the stretch-activated PIEZO1 channel increases the mitosis of cancerous cells in response to tissue stiffness. Similarly, the overexpression of K,1,3 ion channels in tumour specific T-cells helps evacuate potassium ions released by intracellular necrosis which enhances tumour clearance and survival, Evidence is emerging of a positive feedback loop between glioma growth and the increased electrical excitability of the brain tissue surrounding tumours. In Alzheimer disease and related tauopathies/amyloidopathies, tangles of intraneuronal proteins cause brain lesions which modify neuronal function through (i) mechano-electrical coupling to ion channels, (ii) changes in action potential waveforms and (ii) 1WO 20237214156 PCTIGB2023/051155 alterations in calcium and potassium dynamics. Understanding the link between channelopathies and cell function is therefore critical if cognitive functions are to be preserved. There are 850,000 sufferers of dementia in the UK and £1,000 brain tumours are diagnosed each year. These diseases have abysmal prognosis, hence the urgency of finding a quantitative method to diagnose and guide the development of targeted therapies. Currently there is no direct assay that links the electrical function of a cell to the underlying genetic mutations. Genetic sequencing can identify gene mutations but it cannot determine how these mutations affect function. Patch-sequencing methods can correlate, at a qualitative level, gene mutations to morphological characteristics (e.g. axonal arborisation) or the amplitude of action potentials. On the other hand, voltage clamps require multiple pharmacological manipulations and are very time consuming when the affected ion channel is unknown. Further, clinicians cannot perform electrophysiology on healthy human brains celis due to ethical considerations and the paucity of samples. However, healthy assays. are needed as a comparator to diseased cells. Therefore, there is a need for a single shot assay that can infer the fuli complement of ion channels in one go and automatically output all intracellular parameters that underpin electrical waveform. The methods, computer programs and apparatus described in the present disclosure may address one or more of the above issues. Summary According to a first aspect of the present disclosure there is provided a computer- implemented method for determining values of biological parameters of a cell, comprising receiving membrane voltage readings of the cell corresponding to N time points;WO 20237214156 PCTIGB2023/051155 for a state vector comprising the biological parameters and a plurality of interdependent and time-dependent state variables, including 2 membrane voltage variable setting an initial quess of the state vector; performing an optimization of the state vector by: setting the membrane voltage variable to be equal to a corresponding membrane voltage reading at a number of time points within the N time points; determining an updated value of the state vector by optimizing an objective function that operates on values of the membrane voltage variable and corresponding membrane voltage readings; and repeating the optimization with: the initial guess of the state vector set to the updated value of the state vector; and the membrane voltage variable set to be equal to a corresponding membrane voltage reading at fewer time points within the N time points. ‘The cell may be a neuron. The plurality of biological parameters may relate to ‘one or more jon channels of the neuron, The state vector may be of a conductance model of the one or more ion channels of the neuron, ‘The steps of repeating the optimization may be performed a plurality of times. Ina repetition of the optimization, setting the membrane voltage variable to be equal to corresponding membrane voltage reading at fewer time points within the N time points may mean setting the membrane voltage variable at fewer time points than the membrane voltage variable was set in an immediately preceding iteration of the optimization. Depending on the number of times that the optimization has been repeated, the immediately preceding iteration may be the optimization of the state vector in which the membrane voltage variable is set to be equal to the corresponding membrane voltage reading at the number of time points within the N time points, or a subsequent repetition of the optimization. Performing the optimization of the state vector may comprise setting the membrane voltage variable to be equal to a corresponding membrane voltage 3