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Shedding Light on the Blood–Brain Barrier Transport with Two-Photon Microscopy In Vivo

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Abstract

Treatment of brain disorders relies on efficient delivery of therapeutics to the brain, which is hindered by the blood–brain barrier (BBB). The work of Prof. Margareta Hammarlund-Udenaes was instrumental in understanding the principles of drug delivery to the brain and developing new tools to study it. Here, we show how some of the concepts developed in her research can be translated to in vivo 2-photon microscopy (2PM) studies of the BBB. We primarily focus on the methods developed in our laboratory to characterize the paracellular diffusion, adsorptive-mediated transcytosis, and receptor-mediated transcytosis of drug nanocarriers at the microscale, illustrating how 2PM can deepen our understanding of the mechanisms of drug delivery to the brain.

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References

  1. Banks WA. From blood-brain barrier to blood-brain interface: new opportunities for CNS drug delivery. Nat Rev Drug Discov. 2016;15(4):275–92.

    Article  CAS  PubMed  Google Scholar 

  2. Pardridge WM. Drug and gene delivery to the brain: the vascular route. Neuron. 2002;36(4):555–8.

    Article  CAS  PubMed  Google Scholar 

  3. Pardridge WM. Drug transport across the blood-brain barrier. J Cereb Blood Flow Metab. 2012;32(11):1959–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Liddelow S. Bypassing the barrier: new routes for delivery of macromolecules to the central nervous system. J Physiol. 2018;596(3):361–2.

    Article  CAS  PubMed  Google Scholar 

  5. Abbott NJ. Blood-brain barrier structure and function and the challenges for CNS drug delivery. J Inherit Metab Dis. 2013;36(3):437–49.

    Article  CAS  PubMed  Google Scholar 

  6. Fenstermacher JD, Blasberg RG, Patlak CS. Methods for Quantifying the transport of drugs across brain barrier systems. Pharmacol Ther. 1981;14(2):217–48.

    Article  CAS  PubMed  Google Scholar 

  7. Gjedde A. High- and low-affinity transport of D-glucose from blood to brain. J Neurochem. 1981;36(4):1463–71.

    Article  CAS  PubMed  Google Scholar 

  8. Patlak CS, Blasberg RG, Fenstermacher JD. Graphical evaluation of blood-to-brain transfer constants from multiple-time uptake data. JCerebBlood Flow Metab. 1983;3(1):1–7.

    Article  CAS  Google Scholar 

  9. Blasberg RG, Patlak CS, Fenstermacher JD. Selection of experimental conditions for the accurate determination of blood–brain transfer constants from single-time experiments: a theoretical analysis. J Cereb Blood Flow Metab. 1983;3(2):215–25.

    Article  CAS  PubMed  Google Scholar 

  10. Hammarlund-Udenaes M, Paalzow LK, de Lange EC. Drug equilibration across the blood-brain barrier–pharmacokinetic considerations based on the microdialysis method. Pharm Res. 1997;14(2):128–34.

    Article  CAS  PubMed  Google Scholar 

  11. Friden M, Gupta A, Antonsson M, Bredberg U, Hammarlund-Udenaes M. In vitro methods for estimating unbound drug concentrations in the brain interstitial and intracellular fluids. Drug Metab Dispos. 2007;35(9):1711–9.

    Article  CAS  PubMed  Google Scholar 

  12. Loryan I, Hammarlund-Udenaes M, Syvanen S. Brain Distribution of Drugs: Pharmacokinetic Considerations. Handb Exp Pharmacol. 2020.

  13. Chaurasia CS, Muller M, Bashaw ED, Benfeldt E, Bolinder J, Bullock R, Bungay PM, DeLange EC, Derendorf H, Elmquist WF, Hammarlund-Udenaes M, Joukhadar C, Kellogg DL Jr, Lunte CE, Nordstrom CH, Rollema H, Sawchuk RJ, Cheung BW, Shah VP, Stahle L, Ungerstedt U, Welty DF, Yeo H. AAPS-FDA Workshop White Paper: microdialysis principles, application, and regulatory perspectives. J Clin Pharmacol. 2007;47(5):589–603.

    Article  CAS  PubMed  Google Scholar 

  14. Bostrom E, Simonsson US, Hammarlund-Udenaes M. In vivo blood-brain barrier transport of oxycodone in the rat: indications for active influx and implications for pharmacokinetics/pharmacodynamics. Drug Metab Dispos. 2006;34(9):1624–31.

    Article  PubMed  CAS  Google Scholar 

  15. Hammarlund-Udenaes M. Microdialysis as an Important Technique in Systems Pharmacology-a Historical and Methodological Review. AAPS J. 2017;19(5):1294–303.

    Article  CAS  PubMed  Google Scholar 

  16. Friden M, Winiwarter S, Jerndal G, Bengtsson O, Wan H, Bredberg U, Hammarlund-Udenaes M, Antonsson M. Structure-brain exposure relationships in rat and human using a novel data set of unbound drug concentrations in brain interstitial and cerebrospinal fluids. J Med Chem. 2009;52(20):6233–43.

    Article  CAS  PubMed  Google Scholar 

  17. Lindqvist A, Friden M, Hammarlund-Udenaes M. Pharmacokinetic considerations of nanodelivery to the brain: Using modeling and simulations to predict the outcome of liposomal formulations. Eur J Pharm Sci. 2016;92:173–82.

    Article  CAS  PubMed  Google Scholar 

  18. Syvanen S, Xie R, Sahin S, Hammarlund-Udenaes M. Pharmacokinetic consequences of active drug efflux at the blood-brain barrier. Pharm Res. 2006;23(4):705–17.

    Article  CAS  PubMed  Google Scholar 

  19. Friden M, Ljungqvist H, Middleton B, Bredberg U, Hammarlund-Udenaes M. Improved measurement of drug exposure in the brain using drug-specific correction for residual blood. J Cereb Blood Flow Metab. 2010;30(1):150–61.

    Article  CAS  PubMed  Google Scholar 

  20. Friden M, Bergstrom F, Wan H, Rehngren M, Ahlin G, Hammarlund-Udenaes M, Bredberg U. Measurement of unbound drug exposure in brain: modeling of pH partitioning explains diverging results between the brain slice and brain homogenate methods. Drug Metab Dispos. 2011;39(3):353–62.

    Article  CAS  PubMed  Google Scholar 

  21. Gupta A, Chatelain P, Massingham R, Jonsson EN, Hammarlund-Udenaes M. Brain distribution of cetirizine enantiomers: comparison of three different tissue-to-plasma partition coefficients: K(p), K(p, u), and K(p, uu). Drug Metab Dispos. 2006;34(2):318–23.

    Article  CAS  PubMed  Google Scholar 

  22. Loryan I, Sinha V, Mackie C, Van Peer A, Drinkenburg W, Vermeulen A, Morrison D, Monshouwer M, Heald D, Hammarlund-Udenaes M. Mechanistic understanding of brain drug disposition to optimize the selection of potential neurotherapeutics in drug discovery. Pharm Res. 2014;31(8):2203–19.

    Article  CAS  PubMed  Google Scholar 

  23. Luptakova D, Vallianatou T, Nilsson A, Shariatgorji R, Hammarlund-Udenaes M, Loryan I, Andren PE. Neuropharmacokinetic visualization of regional and subregional unbound antipsychotic drug transport across the blood-brain barrier. Mol Psychiatry. 2021.

  24. Lindqvist A, Rip J, Gaillard PJ, Bjorkman S, Hammarlund-Udenaes M. Enhanced brain delivery of the opioid peptide DAMGO in glutathione pegylated liposomes: a microdialysis study. Mol Pharm. 2013;10(5):1533–41.

    Article  CAS  PubMed  Google Scholar 

  25. Hu Y, Gaillard PJ, Rip J, de Lange ECM, Hammarlund-Udenaes M. In Vivo Quantitative Understanding of PEGylated Liposome’s Influence on Brain Delivery of Diphenhydramine. Mol Pharm. 2018;15(12):5493–500.

    Article  CAS  PubMed  Google Scholar 

  26. Ederoth P, Tunblad K, Bouw R, Lundberg CJ, Ungerstedt U, Nordstrom CH, Hammarlund-Udenaes M. Blood-brain barrier transport of morphine in patients with severe brain trauma. Br J Clin Pharmacol. 2004;57(4):427–35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Bengtsson J, Ederoth P, Ley D, Hansson S, Amer-Wahlin I, Hellstrom-Westas L, Marsal K, Nordstrom CH, Hammarlund-Udenaes M. The influence of age on the distribution of morphine and morphine-3-glucuronide across the blood-brain barrier in sheep. Br J Pharmacol. 2009;157(6):1085–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Montagne A, Barnes SR, Sweeney MD, Halliday MR, Sagare AP, Zhao Z, Toga AW, Jacobs RE, Liu CY, Amezcua L, Harrington MG, Chui HC, Law M, Zlokovic BV. Blood-brain barrier breakdown in the aging human hippocampus. Neuron. 2015;85(2):296–302.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Heye AK, Culling RD, Valdes Hernandez Mdel C, Thrippleton MJ, Wardlaw JM. Assessment of blood-brain barrier disruption using dynamic contrast-enhanced MRI. A systematic review. Neuroimage Clin. 2014;6:262–274.

  30. Raja R, Rosenberg GA, Caprihan A. MRI measurements of Blood-Brain Barrier function in dementia: A review of recent studies. Neuropharmacology. 2018;134(Pt B):259–71.

    Article  CAS  PubMed  Google Scholar 

  31. Montagne A, Toga AW, Zlokovic BV. Blood-Brain Barrier Permeability and Gadolinium: Benefits and Potential Pitfalls in Research. JAMA Neurol. 2016;73(1):13–4.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Varatharaj A, Liljeroth M, Darekar A, Larsson HBW, Galea I, Cramer SP. Blood-brain barrier permeability measured using dynamic contrast-enhanced magnetic resonance imaging: a validation study. J Physiol. 2019;597(3):699–709.

    Article  CAS  PubMed  Google Scholar 

  33. van de Haar HJ, Jansen JFA, Jeukens C, Burgmans S, van Buchem MA, Muller M, Hofman PAM, Verhey FRJ, van Osch MJP, Backes WH. Subtle blood-brain barrier leakage rate and spatial extent: Considerations for dynamic contrast-enhanced MRI. Med Phys. 2017;44(8):4112–25.

    Article  PubMed  Google Scholar 

  34. Jelescu IO, Leppert IR, Narayanan S, Araujo D, Arnold DL, Pike GB. Dual-temporal resolution dynamic contrast-enhanced MRI protocol for blood-brain barrier permeability measurement in enhancing multiple sclerosis lesions. J Magn Reson Imaging. 2011;33(6):1291–300.

    Article  CAS  PubMed  Google Scholar 

  35. Verheggen ICM, de Jong JJA, van Boxtel MPJ, Gronenschild E, Palm WM, Postma AA, Jansen JFA, Verhey FRJ, Backes WH. Increase in blood-brain barrier leakage in healthy, older adults. Geroscience. 2020;42(4):1183–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Verheggen ICM, de Jong JJA, van Boxtel MPJ, Postma AA, Jansen JFA, Verhey FRJ, Backes WH. Imaging the role of blood-brain barrier disruption in normal cognitive ageing. Geroscience. 2020;42(6):1751–64.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Cabezon I, Manich G, Martin-Venegas R, Camins A, Pelegri C, Vilaplana J. Trafficking of Gold Nanoparticles Coated with the 8D3 Anti-Transferrin Receptor Antibody at the Mouse Blood-Brain Barrier. Mol Pharm. 2015;12(11):4137–45.

    Article  CAS  PubMed  Google Scholar 

  38. Johnsen KB, Burkhart A, Thomsen LB, Andresen TL, Moos T. Targeting the transferrin receptor for brain drug delivery. Prog Neurobiol. 2019;181:101665.

  39. Allen TM, Cullis PR. Drug delivery systems: entering the mainstream. Science. 2004;303(5665):1818–22.

    Article  CAS  PubMed  Google Scholar 

  40. Couch JA, Yu YJ, Zhang Y, Tarrant JM, Fuji RN, Meilandt WJ, Solanoy H, Tong RK, Hoyte K, Luk W, Lu Y, Gadkar K, Prabhu S, Ordonia BA, Nguyen Q, Lin Y, Lin Z, Balazs M, Scearce-Levie K, Ernst JA, Dennis MS, Watts RJ. Addressing safety liabilities of TfR bispecific antibodies that cross the blood-brain barrier. Sci Transl Med. 2013;5(183):183ra157, 181–112.

  41. Pardridge WM. Blood-Brain Barrier and Delivery of Protein and Gene Therapeutics to Brain. Front Aging Neurosci. 2020;11:373.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  42. Kutuzov N, Flyvbjerg H, Lauritzen M. Contributions of the glycocalyx, endothelium, and extravascular compartment to the blood-brain barrier. Proc Natl Acad Sci U S A. 2018;115(40):E9429–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Lendvai B, Stern EA, Chen B, Svoboda K. Experience-dependent plasticity of dendritic spines in the developing rat barrel cortex in vivo. Nature. 2000;404(6780):876–81.

    Article  CAS  PubMed  Google Scholar 

  44. Helmchen F, Denk W. Deep tissue two-photon microscopy. Nat Methods. 2005;2(12):932–40.

    Article  CAS  PubMed  Google Scholar 

  45. Kucharz K, Kristensen K, Johnsen KB, Lund MA, Lonstrup M, Moos T, Andresen TL, Lauritzen MJ. Post-capillary venules are the key locus for transcytosis-mediated brain delivery of therapeutic nanoparticles. Nat Commun. 2021;12(1):4121.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Engelhardt B, Vajkoczy P, Weller RO. The movers and shapers in immune privilege of the CNS. Nat Immunol. 2017;18(2):123–31.

    Article  CAS  PubMed  Google Scholar 

  47. Thomsen MS, Routhe LJ, Moos T. The vascular basement membrane in the healthy and pathological brain. J Cereb Blood Flow Metab. 2017;37(10):3300–17.

    Article  PubMed  PubMed Central  Google Scholar 

  48. Abbott NJ, Patabendige AA, Dolman DE, Yusof SR, Begley DJ. Structure and function of the blood-brain barrier. Neurobiol Dis. 2010;37(1):13–25.

    Article  CAS  PubMed  Google Scholar 

  49. Tietz S, Engelhardt B. Brain barriers: Crosstalk between complex tight junctions and adherens junctions. J Cell Biol. 2015;209(4):493–506.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Armulik A, Genove G, Mae M, Nisancioglu MH, Wallgard E, Niaudet C, He L, Norlin J, Lindblom P, Strittmatter K, Johansson BR, Betsholtz C. Pericytes regulate the blood-brain barrier. Nature. 2010;468(7323):557–61.

    Article  CAS  PubMed  Google Scholar 

  51. Nhan T, Burgess A, Hynynen K. Transducer design and characterization for dorsal-based ultrasound exposure and two-photon imaging of in vivo blood-brain barrier disruption in a rat model. IEEE Trans Ultrason Ferroelectr Freq Control. 2013;60(7):1376–85.

    Article  PubMed  Google Scholar 

  52. Shi L, Zeng M, Sun Y, Fu BM. Quantification of blood-brain barrier solute permeability and brain transport by multiphoton microscopy. J Biomech Eng. 2014;136(3):031005.

  53. Mathiesen Janiurek M, Soylu-Kucharz R, Christoffersen C, Kucharz K, Lauritzen M. Apolipoprotein M-bound sphingosine-1-phosphate regulates blood-brain barrier paracellular permeability and transcytosis. Elife. 2019;8.

  54. Santisteban MM, Ahn SJ, Lane D, Faraco G, Garcia-Bonilla L, Racchumi G, Poon C, Schaeffer S, Segarra SG, Korbelin J, Anrather J, Iadecola C. Endothelium-Macrophage Crosstalk Mediates Blood-Brain Barrier Dysfunction in Hypertension. Hypertension. 2020;76(3):795–807.

    Article  CAS  PubMed  Google Scholar 

  55. Nikolakopoulou AM, Wang Y, Ma Q, Sagare AP, Montagne A, Huuskonen MT, Rege SV, Kisler K, Dai Z, Korbelin J, Herz J, Zhao Z, Zlokovic BV. Endothelial LRP1 protects against neurodegeneration by blocking cyclophilin A. J Exp Med. 2021;218(4).

  56. Lee S, Kang BM, Kim JH, Min J, Kim HS, Ryu H, Park H, Bae S, Oh D, Choi M, Suh M. Real-time in vivo two-photon imaging study reveals decreased cerebro-vascular volume and increased blood-brain barrier permeability in chronically stressed mice. Sci Rep. 2018;8(1):13064.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  57. Kucharz K, Sondergaard Rasmussen I, Bach A, Stromgaard K, Lauritzen M. PSD-95 uncoupling from NMDA receptors by Tat-N-dimer ameliorates neuronal depolarisation in cortical spreading depression. J Cereb Blood Flow Metab. 2016.

  58. Nhan T, Burgess A, Cho EE, Stefanovic B, Lilge L, Hynynen K. Drug delivery to the brain by focused ultrasound induced blood-brain barrier disruption: quantitative evaluation of enhanced permeability of cerebral vasculature using two-photon microscopy. J Control Release. 2013;172(1):274–80.

    Article  CAS  PubMed  Google Scholar 

  59. Verant P, Serduc R, van der Sanden B, Chantal R, Ricard C, Coles JA, Vial JC. Subtraction method for intravital two-photon microscopy: intraparenchymal imaging and quantification of extravasation in mouse brain cortex. J Biomed Opt. 2008;13(1):011002.

  60. Hammarlund-Udenaes M, Friden M, Syvanen S, Gupta A. On the rate and extent of drug delivery to the brain. Pharm Res. 2008;25(8):1737–50.

    Article  CAS  PubMed  Google Scholar 

  61. Abbott NJ, Patabendige AA, Dolman DE, Yusof SR, Begley DJ. Structure and function of the blood-brain barrier. NeurobiolDis. 2010;37(1):13–25.

    CAS  Google Scholar 

  62. Richard JP, Melikov K, Vives E, Ramos C, Verbeure B, Gait MJ, Chernomordik LV, Lebleu B. Cell-penetrating peptides. A reevaluation of the mechanism of cellular uptake. J Biol Chem. 2003;278(1):585–590.

  63. Madani F, Lindberg S, Langel U, Futaki S, Graslund A. Mechanisms of cellular uptake of cell-penetrating peptides. J Biophys. 2011;2011:414729.

  64. Trabulo S, Cardoso AL, Mano M, De Lima MC. Cell-Penetrating Peptides-Mechanisms of Cellular Uptake and Generation of Delivery Systems. Pharmaceuticals (Basel). 2010;3(4):961–93.

    Article  CAS  Google Scholar 

  65. Guo Z, Peng H, Kang J, Sun D. Cell-penetrating peptides: Possible transduction mechanisms and therapeutic applications. Biomed Rep. 2016;4(5):528–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Rizzuti M, Nizzardo M, Zanetta C, Ramirez A, Corti S. Therapeutic applications of the cell-penetrating HIV-1 Tat peptide. Drug Discov Today. 2015;20(1):76–85.

    Article  CAS  PubMed  Google Scholar 

  67. Froslev P, Franzyk H, Ozgur B, Brodin B, Kristensen M. Highly cationic cell-penetrating peptides affect the barrier integrity and facilitates mannitol permeation in a human stem cell-based blood-brain barrier model. Eur J Pharm Sci. 2022;168:106054.

  68. Gabathuler R. Approaches to transport therapeutic drugs across the blood-brain barrier to treat brain diseases. Neurobiol Dis. 2010;37(1):48–57.

    Article  CAS  PubMed  Google Scholar 

  69. Farkhani SM, Valizadeh A, Karami H, Mohammadi S, Sohrabi N, Badrzadeh F. Cell penetrating peptides: efficient vectors for delivery of nanoparticles, nanocarriers, therapeutic and diagnostic molecules. Peptides. 2014;57:78–94.

    Article  CAS  PubMed  Google Scholar 

  70. Bach A, Clausen BH, Moller M, Vestergaard B, Chi CN, Round A, Sorensen PL, Nissen KB, Kastrup JS, Gajhede M, Jemth P, Kristensen AS, Lundstrom P, Lambertsen KL, Stromgaard K. A high-affinity, dimeric inhibitor of PSD-95 bivalently interacts with PDZ1-2 and protects against ischemic brain damage. Proc Natl Acad Sci U S A. 2012;109(9):3317–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. Kucharz K, Sondergaard Rasmussen I, Bach A, Stromgaard K, Lauritzen M. PSD-95 uncoupling from NMDA receptors by Tat- N-dimer ameliorates neuronal depolarization in cortical spreading depression. J Cereb Blood Flow Metab. 2017;37(5):1820–8.

    Article  CAS  PubMed  Google Scholar 

  72. Dreier JP, Fabricius M, Ayata C, Sakowitz OW, William Shuttleworth C, Dohmen C, Graf R, Vajkoczy P, Helbok R, Suzuki M, Schiefecker AJ, Major S, Winkler MK, Kang EJ, Milakara D, Oliveira-Ferreira AI, Reiffurth C, Revankar GS, Sugimoto K, Dengler NF, Hecht N, Foreman B, Feyen B, Kondziella D, Friberg CK, Piilgaard H, Rosenthal ES, Westover MB, Maslarova A, Santos E, Hertle D, Sanchez-Porras R, Jewell SL, Balanca B, Platz J, Hinzman JM, Luckl J, Schoknecht K, Scholl M, Drenckhahn C, Feuerstein D, Eriksen N, Horst V, Bretz JS, Jahnke P, Scheel M, Bohner G, Rostrup E, Pakkenberg B, Heinemann U, Claassen J, Carlson AP, Kowoll CM, Lublinsky S, Chassidim Y, Shelef I, Friedman A, Brinker G, Reiner M, Kirov SA, Andrew RD, Farkas E, Guresir E, Vatter H, Chung LS, Brennan KC, Lieutaud T, Marinesco S, Maas AI, Sahuquillo J, Dahlem MA, Richter F, Herreras O, Boutelle MG, Okonkwo DO, Bullock MR, Witte OW, Martus P, van den Maagdenberg AM, Ferrari MD, Dijkhuizen RM, Shutter LA, Andaluz N, Schulte AP, MacVicar B, Watanabe T, Woitzik J, Lauritzen M, Strong AJ, Hartings JA. Recording, analysis, and interpretation of spreading depolarizations in neurointensive care: Review and recommendations of the COSBID research group. J Cereb Blood Flow Metab. 2017;37(5):1595–625.

    Article  PubMed  Google Scholar 

  73. Hartings JA, Shuttleworth CW, Kirov SA, Ayata C, Hinzman JM, Foreman B, Andrew RD, Boutelle MG, Brennan KC, Carlson AP, Dahlem MA, Drenckhahn C, Dohmen C, Fabricius M, Farkas E, Feuerstein D, Graf R, Helbok R, Lauritzen M, Major S, Oliveira-Ferreira AI, Richter F, Rosenthal ES, Sakowitz OW, Sanchez-Porras R, Santos E, Scholl M, Strong AJ, Urbach A, Westover MB, Winkler MK, Witte OW, Woitzik J, Dreier JP. The continuum of spreading depolarizations in acute cortical lesion development: Examining Leao’s legacy. J Cereb Blood Flow Metab. 2017;37(5):1571–94.

    Article  PubMed  Google Scholar 

  74. Dreier JP, Reiffurth C. The stroke-migraine depolarization continuum. Neuron. 2015;86(4):902–22.

    Article  CAS  PubMed  Google Scholar 

  75. Lauritzen M, Dreier JP, Fabricius M, Hartings JA, Graf R, Strong AJ. Clinical relevance of cortical spreading depression in neurological disorders: migraine, malignant stroke, subarachnoid and intracranial hemorrhage, and traumatic brain injury. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism. 2011;31(1):17–35.

    Article  Google Scholar 

  76. Tunnemann G, Martin RM, Haupt S, Patsch C, Edenhofer F, Cardoso MC. Cargo-dependent mode of uptake and bioavailability of TAT-containing proteins and peptides in living cells. FASEB J. 2006;20(11):1775–84.

    Article  PubMed  CAS  Google Scholar 

  77. Kristensen M, Kucharz K, Felipe Alves Fernandes E, Stromgaard K, Schallburg Nielsen M, Cederberg Helms HC, Bach A, Ulrikkaholm Tofte-Hansen M, Irene Aldana Garcia B, Lauritzen M, Brodin B. Conjugation of Therapeutic PSD-95 Inhibitors to the Cell-Penetrating Peptide Tat Affects Blood-Brain Barrier Adherence, Uptake, and Permeation. Pharmaceutics. 2020;12(7).

  78. Loryan I, Melander E, Svensson M, Payan M, König F, Jansson B, Hammarlund-Udenaes M. In-depth neuropharmacokinetic analysis of antipsychotics based on a novel approach to estimate unbound target-site concentration in CNS regions: link to spatial receptor occupancy. Mol Psychiatry. 2016;21(11):1527–36.

    Article  CAS  PubMed  Google Scholar 

  79. Uusna J, Langel K, Langel U. Toxicity, Immunogenicity, Uptake, and Kinetics Methods for CPPs. Methods Mol Biol. 2015;1324:133–48.

    Article  PubMed  Google Scholar 

  80. De Bock M, Van Haver V, Vandenbroucke RE, Decrock E, Wang N, Leybaert L. Into rather unexplored terrain-transcellular transport across the blood-brain barrier. Glia. 2016;64(7):1097–123.

    Article  PubMed  Google Scholar 

  81. Villasenor R, Lampe J, Schwaninger M, Collin L. Intracellular transport and regulation of transcytosis across the blood-brain barrier. Cell Mol Life Sci. 2019;76(6):1081–92.

    Article  CAS  PubMed  Google Scholar 

  82. Ben-Zvi A, Lacoste B, Kur E, Andreone BJ, Mayshar Y, Yan H, Gu C. Mfsd2a is critical for the formation and function of the blood-brain barrier. Nature. 2014;509(7501):507–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Andreone BJ, Chow BW, Tata A, Lacoste B, Ben-Zvi A, Bullock K, Deik AA, Ginty DD, Clish CB, Gu C. Blood-Brain Barrier Permeability Is Regulated by Lipid Transport-Dependent Suppression of Caveolae-Mediated Transcytosis. Neuron. 2017;94(3):581–594 e585.

  84. Abbott NJ. Astrocyte-endothelial interactions and blood-brain barrier permeability. J Anat. 2002;200(6):629–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Knowland D, Arac A, Sekiguchi KJ, Hsu M, Lutz SE, Perrino J, Steinberg GK, Barres BA, Nimmerjahn A, Agalliu D. Stepwise recruitment of transcellular and paracellular pathways underlies blood-brain barrier breakdown in stroke. Neuron. 2014;82(3):603–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. Habgood MD, Bye N, Dziegielewska KM, Ek CJ, Lane MA, Potter A, Morganti-Kossmann C, Saunders NR. Changes in blood-brain barrier permeability to large and small molecules following traumatic brain injury in mice. Eur J Neurosci. 2007;25(1):231–8.

    Article  CAS  PubMed  Google Scholar 

  87. Yang AC, Stevens MY, Chen MB, Lee DP, Stahli D, Gate D, Contrepois K, Chen W, Iram T, Zhang L, Vest RT, Chaney A, Lehallier B, Olsson N, du Bois H, Hsieh R, Cropper HC, Berdnik D, Li L, Wang EY, Traber GM, Bertozzi CR, Luo J, Snyder MP, Elias JE, Quake SR, James ML, Wyss-Coray T. Physiological blood-brain transport is impaired with age by a shift in transcytosis. Nature. 2020.

  88. Sadeghian H, Lacoste B, Qin T, Toussay X, Rosa R, Oka F, Chung DY, Takizawa T, Gu C, Ayata C. Spreading depolarizations trigger caveolin-1-dependent endothelial transcytosis. Ann Neurol. 2018;84(3):409–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  89. Montagne A, Zhao Z, Zlokovic BV. Alzheimer’s disease: A matter of blood-brain barrier dysfunction? J Exp Med. 2017;214(11):3151–69.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  90. Sorets AG, Rosch JC, Duvall CL, Lippmann ES. Caveolae-Mediated Transport at the Injured Blood-Brain Barrier as an Underexplored Pathway for Central Nervous System Drug Delivery. Curr Opin Chem Eng. 2020;30:86–95.

    Article  PubMed  PubMed Central  Google Scholar 

  91. Herve F, Ghinea N, Scherrmann JM. CNS delivery via adsorptive transcytosis. AAPS J. 2008;10(3):455–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  92. Gaillard PJ, Visser CC, de Boer M, Appeldoorn CCM, Rip J. Blood-to-Brain Drug Delivery Using Nanocarriers. In: Hammarlund-Udenaes M, de Lange ECM, Thorne RG, editors. Drug Delivery to the Brain: Physiological Concepts, Methodologies and Approaches. New York, NY: Springer New York; 2014. p. 433–454.

  93. Larsen MT, Kuhlmann M, Hvam ML, Howard KA. Albumin-based drug delivery: harnessing nature to cure disease. Mol Cell Ther. 2016;4:3–3.

    Article  PubMed  PubMed Central  Google Scholar 

  94. Sleep D. Albumin and its application in drug delivery. Expert Opin Drug Deliv. 2015;12(5):793–812.

    Article  CAS  PubMed  Google Scholar 

  95. Pilati D, Howard KA. Albumin-based drug designs for pharmacokinetic modulation. Expert Opin Drug Metab Toxicol. 2020;16(9):783–95.

    Article  CAS  PubMed  Google Scholar 

  96. Tuma P, Hubbard AL. Transcytosis: crossing cellular barriers. Physiol Rev. 2003;83(3):871–932.

    Article  CAS  PubMed  Google Scholar 

  97. Sharma HS, Olsson Y, Dey PK. Changes in blood-brain barrier and cerebral blood flow following elevation of circulating serotonin level in anesthetized rats. Brain Res. 1990;517:215–23.

    Article  CAS  PubMed  Google Scholar 

  98. Janiurek M, Soylu-Kucharz R, Christoffersen C, Kucharz K, Lauritzen M. Apolipoprotein M-bound sphingosine-1-phosphate regulates blood-brain barrier paracellular permeability and transcytosis. Elife. 2019;8.

  99. Christensen PM, Liu CH, Swendeman SL, Obinata H, Qvortrup K, Nielsen LB, Hla T, Di Lorenzo A, Christoffersen C. Impaired endothelial barrier function in apolipoprotein M-deficient mice is dependent on sphingosine-1-phosphate receptor 1. FASEB J. 2016;30(6):2351–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  100. Christoffersen C, Obinata H, Kumaraswamy SB, Galvani S, Ahnstrom J, Sevvana M, Egerer-Sieber C, Muller YA, Hla T, Nielsen LB, Dahlback B. Endothelium-protective sphingosine-1-phosphate provided by HDL-associated apolipoprotein M. Proc Natl Acad Sci U S A. 2011;108(23):9613–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  101. Lee MJ, Van Brocklyn JR, Thangada S, Liu CH, Hand AR, Menzeleev R, Spiegel S, Hla T. Sphingosine-1-phosphate as a ligand for the G protein-coupled receptor EDG-1. Science. 1998;279(5356):1552–5.

    Article  CAS  PubMed  Google Scholar 

  102. Yanagida K, Liu CH, Faraco G, Galvani S, Smith HK, Burg N, Anrather J, Sanchez T, Iadecola C, Hla T. Size-selective opening of the blood-brain barrier by targeting endothelial sphingosine 1-phosphate receptor 1. Proc Natl Acad Sci U S A. 2017;114(17):4531–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  103. Saunders NR, Dziegielewska KM, Mollgard K, Habgood MD. Physiology and molecular biology of barrier mechanisms in the fetal and neonatal brain. J Physiol. 2018;596(23):5723–56.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  104. Vanlandewijck M, He L, Mae MA, Andrae J, Ando K, Del Gaudio F, Nahar K, Lebouvier T, Lavina B, Gouveia L, Sun Y, Raschperger E, Rasanen M, Zarb Y, Mochizuki N, Keller A, Lendahl U, Betsholtz C. A molecular atlas of cell types and zonation in the brain vasculature. Nature. 2018;554(7693):475–80.

    Article  CAS  PubMed  Google Scholar 

  105. Wilhelm I, Nyul-Toth A, Suciu M, Hermenean A, Krizbai IA. Heterogeneity of the blood-brain barrier. Tissue Barriers. 2016;4(1):e1143544.

  106. Lu W. Adsorptive-mediated brain delivery systems. Curr Pharm Biotechnol. 2012;13(12):2340–8.

    Article  CAS  PubMed  Google Scholar 

  107. Pulgar VM. Transcytosis to Cross the Blood Brain Barrier. New Advancements and Challenges Front Neurosci. 2018;12:1019.

    Article  PubMed  Google Scholar 

  108. Pardridge WM. The blood-brain barrier and neurotherapeutics. NeuroRx. 2005;2(1):1–2.

    Article  PubMed  PubMed Central  Google Scholar 

  109. Pardridge WM. The blood-brain barrier: bottleneck in brain drug development. NeuroRx. 2005;2(1):3–14.

    Article  PubMed  PubMed Central  Google Scholar 

  110. Sercombe L, Veerati T, Moheimani F, Wu SY, Sood AK, Hua S. Advances and Challenges of Liposome Assisted Drug Delivery. Front Pharmacol. 2015;6:286.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  111. Rip J. Liposome technologies and drug delivery to the CNS. Drug Discov Today Technol. 2016;20:53–8.

    Article  PubMed  Google Scholar 

  112. Lai F, Fadda AM, Sinico C. Liposomes for brain delivery. Expert Opin Drug Deliv. 2013;10(7):1003–22.

    Article  CAS  PubMed  Google Scholar 

  113. Hu Y, Hammarlund-Udenaes M. Perspectives on Nanodelivery to the Brain: Prerequisites for Successful Brain Treatment. Mol Pharm. 2020;17(11):4029–39.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  114. Jefferies WA, Brandon MR, Hunt SV, Williams AF, Gatter KC, Mason DY. Transferrin receptor on endothelium of brain capillaries. Nature. 1984;312(5990):162–3.

    Article  CAS  PubMed  Google Scholar 

  115. Moos T, Oates PS, Morgan EH. Expression of the neuronal transferrin receptor is age dependent and susceptible to iron deficiency. J Comp Neurol. 1998;398(3):420–30.

    Article  CAS  PubMed  Google Scholar 

  116. Niewoehner J, Bohrmann B, Collin L, Urich E, Sade H, Maier P, Rueger P, Stracke JO, Lau W, Tissot AC, Loetscher H, Ghosh A, Freskgard PO. Increased brain penetration and potency of a therapeutic antibody using a monovalent molecular shuttle. Neuron. 2014;81(1):49–60.

    Article  CAS  PubMed  Google Scholar 

  117. Yu YJ, Zhang Y, Kenrick M, Hoyte K, Luk W, Lu Y, Atwal J, Elliott JM, Prabhu S, Watts RJ, Dennis MS. Boosting brain uptake of a therapeutic antibody by reducing its affinity for a transcytosis target. Sci Transl Med. 2011;3(84):84ra44.

  118. Ullman JC, Arguello A, Getz JA, Bhalla A, Mahon CS, Wang J, Giese T, Bedard C, Kim DJ, Blumenfeld JR, Liang N, Ravi R, Nugent AA, Davis SS, Ha C, Duque J, Tran HL, Wells RC, Lianoglou S, Daryani VM, Kwan W, Solanoy H, Nguyen H, Earr T, Dugas JC, Tuck MD, Harvey JL, Reyzer ML, Caprioli RM, Hall S, Poda S, Sanchez PE, Dennis MS, Gunasekaran K, Srivastava A, Sandmann T, Henne KR, Thorne RG, Di Paolo G, Astarita G, Diaz D, Silverman AP, Watts RJ, Sweeney ZK, Kariolis MS, Henry AG. Brain delivery and activity of a lysosomal enzyme using a blood-brain barrier transport vehicle in mice. Sci Transl Med. 2020;12(545).

  119. Kariolis MS, Wells RC, Getz JA, Kwan W, Mahon CS, Tong R, Kim DJ, Srivastava A, Bedard C, Henne KR, Giese T, Assimon VA, Chen X, Zhang Y, Solanoy H, Jenkins K, Sanchez PE, Kane L, Miyamoto T, Chew KS, Pizzo ME, Liang N, Calvert MEK, DeVos SL, Baskaran S, Hall S, Sweeney ZK, Thorne RG, Watts RJ, Dennis MS, Silverman AP, Zuchero YJY. Brain delivery of therapeutic proteins using an Fc fragment blood-brain barrier transport vehicle in mice and monkeys. Sci Transl Med. 2020;12(545).

  120. Lee HJ, Engelhardt B, Lesley J, Bickel U, Pardridge WM. Targeting rat anti-mouse transferrin receptor monoclonal antibodies through blood-brain barrier in mouse. J Pharmacol Exp Ther. 2000;292(3):1048–52.

    CAS  PubMed  Google Scholar 

  121. Johnsen KB, Bak M, Melander F, Thomsen MS, Burkhart A, Kempen PJ, Andresen TL, Moos T. Modulating the antibody density changes the uptake and transport at the blood-brain barrier of both transferrin receptor-targeted gold nanoparticles and liposomal cargo. J Control Release. 2019;295:237–49.

    Article  CAS  PubMed  Google Scholar 

  122. Lajoie JM, Shusta EV. Targeting receptor-mediated transport for delivery of biologics across the blood-brain barrier. Annu Rev Pharmacol Toxicol. 2015;55:613–31.

    Article  CAS  PubMed  Google Scholar 

  123. Barenholz Y. Doxil(R)–the first FDA-approved nano-drug: lessons learned. J Control Release. 2012;160(2):117–34.

    Article  CAS  PubMed  Google Scholar 

  124. Johnsen KB, Burkhart A, Melander F, Kempen PJ, Vejlebo JB, Siupka P, Nielsen MS, Andresen TL, Moos T. Targeting transferrin receptors at the blood-brain barrier improves the uptake of immunoliposomes and subsequent cargo transport into the brain parenchyma. Sci Rep. 2017;7(1):10396.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  125. Hadaczek P, Yamashita Y, Mirek H, Tamas L, Bohn MC, Noble C, Park JW, Bankiewicz K. The, “perivascular pump” driven by arterial pulsation is a powerful mechanism for the distribution of therapeutic molecules within the brain. Mol Ther. 2006;14(1):69–78.

    Article  CAS  PubMed  Google Scholar 

  126. Mestre H, Tithof J, Du T, Song W, Peng W, Sweeney AM, Olveda G, Thomas JH, Nedergaard M, Kelley DH. Flow of cerebrospinal fluid is driven by arterial pulsations and is reduced in hypertension. Nat Commun. 2018;9(1):4878.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  127. Zhang ET, Inman CB, Weller RO. Interrelationships of the pia mater and the perivascular (Virchow-Robin) spaces in the human cerebrum. J Anat. 1990;170:111–23.

    CAS  PubMed  PubMed Central  Google Scholar 

  128. Owens T, Bechmann I, Engelhardt B. Perivascular spaces and the two steps to neuroinflammation. J Neuropathol Exp Neurol. 2008;67(12):1113–21.

    Article  PubMed  Google Scholar 

  129. Wardlaw JM, Benveniste H, Nedergaard M, Zlokovic BV, Mestre H, Lee H, Doubal FN, Brown R, Ramirez J, MacIntosh BJ, Tannenbaum A, Ballerini L, Rungta RL, Boido D, Sweeney M, Montagne A, Charpak S, Joutel A, Smith KJ, Black SE, colleagues from the Fondation Leducq Transatlantic Network of Excellence on the Role of the Perivascular Space in Cerebral Small Vessel D. Perivascular spaces in the brain: anatomy, physiology and pathology. Nat Rev Neurol. 2020;16(3):137–153.

  130. Foley CP, Nishimura N, Neeves KB, Schaffer CB, Olbricht WL. Real-time imaging of perivascular transport of nanoparticles during convection-enhanced delivery in the rat cortex. Ann Biomed Eng. 2012;40(2):292–303.

    Article  PubMed  Google Scholar 

  131. Muldoon LL, Pagel MA, Kroll RA, Roman-Goldstein S, Jones RS, Neuwelt EA. A physiological barrier distal to the anatomic blood-brain barrier in a model of transvascular delivery. AJNR Am J Neuroradiol. 1999;20(2):217–22.

    CAS  PubMed  PubMed Central  Google Scholar 

  132. Thorne RG, Nicholson C. In vivo diffusion analysis with quantum dots and dextrans predicts the width of brain extracellular space. Proc Natl Acad Sci U S A. 2006;103(14):5567–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  133. Tonnesen J, Inavalli V, Nagerl UV. Super-Resolution Imaging of the Extracellular Space in Living Brain Tissue. Cell. 2018;172(5):1108–1121 e1115.

  134. Nance EA, Woodworth GF, Sailor KA, Shih TY, Xu Q, Swaminathan G, Xiang D, Eberhart C, Hanes J. A dense poly(ethylene glycol) coating improves penetration of large polymeric nanoparticles within brain tissue. Sci Transl Med. 2012;4(149):149ra119.

  135. Aggarwal P, Hall JB, McLeland CB, Dobrovolskaia MA, McNeil SE. Nanoparticle interaction with plasma proteins as it relates to particle biodistribution, biocompatibility and therapeutic efficacy. Adv Drug Deliv Rev. 2009;61(6):428–37.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  136. Lynch I, Cedervall T, Lundqvist M, Cabaleiro-Lago C, Linse S, Dawson KA. The nanoparticle-protein complex as a biological entity; a complex fluids and surface science challenge for the 21st century. Adv Colloid Interface Sci. 2007;134–135:167–74.

    Article  PubMed  CAS  Google Scholar 

  137. Rampado R, Crotti S, Caliceti P, Pucciarelli S, Agostini M. Recent Advances in Understanding the Protein Corona of Nanoparticles and in the Formulation of “Stealthy” Nanomaterials. Front Bioeng Biotechnol. 2020;8:166.

    Article  PubMed  PubMed Central  Google Scholar 

  138. Suk JS, Xu Q, Kim N, Hanes J, Ensign LM. PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Adv Drug Deliv Rev. 2016;99(Pt A):28–51.

    Article  CAS  PubMed  Google Scholar 

  139. Verhoef JJ, Anchordoquy TJ. Questioning the Use of PEGylation for Drug Delivery. Drug Deliv Transl Res. 2013;3(6):499–503.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  140. Jing Y, Zhang C, Yu B, Lin D, Qu J. Super-Resolution Microscopy: Shedding New Light on In Vivo Imaging. Front Chem. 2021;9:746900.

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All authors declare no conflict of interest.

Funding

This work was supported by the Læge Sofus Carl Emil Friis og Hustru Olga Doris Friis’ Legat; the Lundbeck Foundation; the Danish Medical Research Council; the NOVO Nordisk Foundation, and a Nordea Foundation grant to the Center for Healthy Aging.

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  1. Department of Neuroscience, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark

    Krzysztof Kucharz, Nikolay Kutuzov, Oleg Zhukov, Mette Mathiesen Janiurek & Martin Lauritzen

  2. Department of Clinical Neurophysiology, Rigshospitalet, Glostrup, Denmark

    Martin Lauritzen

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  1. Krzysztof Kucharz
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Kucharz, K., Kutuzov, N., Zhukov, O. et al. Shedding Light on the Blood–Brain Barrier Transport with Two-Photon Microscopy In Vivo. Pharm Res 39, 1457–1468 (2022). https://doi.org/10.1007/s11095-022-03266-2

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