Improving brain structure and biomarkers in alzheimer's disease with mesenchymal stem cells

Abstract

Disclosed herein are compositions and methods for treating Alzheimer's disease with allogeneic mesenchymal stem cells. The treatment methods comprise administering a composition of allogeneic mesenchymal stem cells to a subject in need thereof, wherein the efficacy of the treatment methods can be determined by measuring certain biomarkers and improved cognitive function and / or quality of life.

Description

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 612,240, filed December 19, 2023, which is incorporated herein by reference as if it had been fully rewritten herein.

[0002] field

[0003] This application relates to methods and compositions for treating Alzheimer's disease in subjects in need. Some embodiments involve compositions comprising a therapeutically effective amount of allogeneic mesenchymal stem cells (MSCs) for alleviating symptoms of Alzheimer's disease. Other embodiments involve treatment methods in which a composition comprising a therapeutically effective amount of MSCs is administered to a subject suffering from symptoms of Alzheimer's disease. The effectiveness of these treatments is assessed by measuring specific biomarkers in the subject after administration of the MSC-containing composition, examining changes in their brain activity or morphology, and determining whether their cognitive function or quality of life improves after treatment.

[0004] background

[0005] Alzheimer's disease (AD) involves a complex pathology and encompasses multiple mechanisms beyond β-amyloid deposition and neurofibrillary tangles. It is increasingly recognized that pro-inflammatory states contribute to subsequent dementia. In this regard, pro-inflammatory cytokines are abundant near amyloid deposits and neurofibrillary tangles, and there is a link between systemic inflammation and β-amyloid accumulation. AD is also characterized by neurovascular damage leading to adverse outcomes. The resulting blood-brain barrier (BBB) ​​impairment impairs transendothelial exchange, resulting in inefficient clearance and accumulation of AβP in the brain.

[0006] Due to the complex nature of Alzheimer's disease (AD) progression, using biomarkers to predict AD onset and progression remains challenging. While the concentrations of β-amyloid deposits and neurofibrillary tangles can be used to diagnose or predict AD onset, some individuals exhibit significant amyloid deposits and neurofibrillary tangles at autopsy, qualifying them for an AD diagnosis despite never having a history of dementia. Currently approved AD treatments (Rivastigmine, Donepezil, Memantine, Galantamine, and Tacrine) offer only marginal benefits and are primarily symptomatic; no approved therapies effectively stop, reverse, or prevent AD. The continued failure of initially promising lead compounds has resulted in no new AD drugs being approved for over a decade. Recent failures include the anti-amyloid monoclonal antibodies solanezumab (Ely Lily) and aducanumab (Biogen / Eisai), which were found to be ineffective against mild to moderate stages of Alzheimer's disease (AD) and mild cognitive impairment (MCI). A common theme among these failures is their targeting of a single pathological feature in AD.

[0007] Simultaneously addressing these neuropathological features of Alzheimer's disease (AD) can offer therapeutic advantages and generate new treatment strategies. Medicinal signaling cells (MSCs, also known as mesenchymal stem cells) are pluripotent cells (in vitro) with pleiotropic mechanisms of action (MOA), including anti-inflammatory properties, the ability to improve vascular function, and the promotion of intrinsic tissue repair and regeneration. MSCs are transported to sites of inflammation and injury, and can therefore target neuroinflammatory sites in AD. MSCs can also modulate the host stem cell niche through paracrine activity and xenocellular coupling to promote intrinsic repair and regeneration. Finally, MSCs are immune-evading / immune-exempt, allowing for allogeneic use, and have an acceptable safety profile in clinical trials. These immune-exempt / immune-evading properties, due to their undetectable levels of major histocompatibility complex II (MHC-II) molecules and low MHC-I levels, make mesenchymal stem cells a potential “off-the-shelf” therapy that is readily available and usable by a broad patient population.

[0008] Preclinical data support the efficacy of MSCs in Alzheimer's disease (AD). In animal models, MSCs cross the brain-brain barrier (BBB), promote neurogenesis, inhibit β-amyloid deposition and promote clearance, reduce apoptosis, promote hippocampal neurogenesis, improve dendritic morphology, and enhance behavioral and spatial memory. These beneficial effects are associated with reduced inflammation, increased Aβ degradation factors and Aβ clearance, reduced hyperphosphorylated tau protein, and elevated markers of alternative activated microglia. These benefits are at least partly due to Aβ-induced release of chemokines from MSCs, which recruit alternative microglia to the brain to reduce Aβ deposition. MSCs have been reported to be effective in young AD model mice prior to Aβ accumulation, resulting in a significant reduction in brain Aβ deposition and a significant increase in presynaptic protein expression. Impressively, these effects lasted for at least 2 months, suggesting that MSCs may be effective as an interventional therapeutic agent for pre-AD.

[0009] Therefore, this application seeks not only to provide a method for treating AD, wherein the method includes using a composition containing MSCs, but also to provide a method for evaluating its efficacy in treating AD in subjects in need and in alleviating AD symptoms. Summary of the Invention

[0010] The purpose of this application is to provide biomarkers indicating efficacy in alleviating AD symptoms or treating Alzheimer's disease (AD) or inhibiting the progression of AD in subjects with symptoms of Alzheimer's disease (AD), wherein one or more biomarkers in the subject are measured before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). These biomarkers may be size changes in brain regions of the patient, such as the whole brain, lateral ventricles, gray matter, hippocampus, temporal cortex, medial temporal cortex, hippocampus, thalamus, white matter, cingulate cortex, and / or frontal cortex. Other biomarkers may be changes in diffusion rate in the cingulate cortex, and / or changes in one or more of whole blood, plasma, or serum biomarkers, such as, but not limited to, levels of soluble TIE2 (sTIE2), eosinophil chemokine 1, eosinophil chemokine 2, or eosinophil chemokine 3.

[0011] In some implementations, the biomarker is an MRI biomarker.

[0012] In some embodiments, the method includes measuring the cognitive function of subjects with AD symptoms before and after administration of a composition containing allogeneic MSCs.

[0013] In some embodiments, the method includes assessing the quality of life of subjects with AD symptoms before and after administration of a composition containing allogeneic MSCs.

[0014] In some implementations, MRI biomarkers include volume measurements of the whole brain, lateral ventricles, gray matter, hippocampus, temporal cortex, medial temporal cortex, hippocampus, thalamus, white matter, cingulate cortex, frontal cortex, or a combination thereof.

[0015] In some embodiments, MRI biomarkers may be a reduction in the volume of the whole brain, gray matter, temporal lobe cortex, medial temporal lobe cortex, hippocampus, frontal cortex, or thalamus. In a preferred embodiment, the degree of reduction in the volume of the whole brain, gray matter, temporal lobe cortex, medial temporal lobe cortex, hippocampus, frontal cortex, or thalamus after administration of allogeneic MSCs to the subject is an indicator of efficacy. The reduction in volume may be 1% to 5%, 5% to 10%, 10% to 50%, or greater than 50%.

[0016] In some implementations, the MRI biomarker may be an increase in the volume of the lateral ventricle, white matter, or cingulate cortex. In a preferred implementation, the decrease in the degree of increase in the volume of the lateral ventricle, white matter, or cingulate cortex after administration of allogeneic MSCs to the subject is an indicator of efficacy. The volume reduction may be 1% to 5%, 5% to 10%, 10% to 50%, or greater than 50%.

[0017] In some embodiments, MRI biomarkers may be mean diffusion rate or free water measurements via diffusion tensor imaging (DTI) or similar techniques in the cingulate cortex. In a preferred embodiment, a decrease in mean diffusion rate or free water measurements in the cingulate cortex via DTI or similar techniques after administration of allogeneic MSCs to the subject is an indication of efficacy.

[0018] In some embodiments, the biomarker may be a whole blood, plasma, or serum biomarker, or one or more of multiple biomarkers. In some embodiments, the serum or plasma biomarker is the level of soluble TIE2 (sTIE2). In a preferred embodiment, the level of sTIE2 in serum or plasma decreases after administration of allogeneic MSCs to the subject.

[0019] In some embodiments, a composition comprising allogeneic mesenchymal stem cells (MSCs) is administered to alleviate symptoms of Alzheimer's disease (AD), treat AD, and / or inhibit the progression of AD, said composition comprising 25 × 10⁻⁶ MSCs. 6 MSC.

[0020] In other embodiments, a composition comprising allogeneic mesenchymal stem cells (MSCs) is administered to alleviate AD symptoms, treat AD, and / or inhibit AD disease progression, said composition comprising 100 × 10 6 MSC.

[0021] In some implementations, a composition comprising allogeneic mesenchymal stem cells (MSCs) is administered to a subject via intravenous or intra-arterial infusion to alleviate AD symptoms, treat AD, and / or inhibit the progression of AD disease.

[0022] In some embodiments, a composition comprising allogeneic mesenchymal stem cells (MSCs) is administered to alleviate AD symptoms, treat AD, and / or inhibit AD disease progression, said composition being administered as one or more of a single dose, a monthly dose, and a repeated interval dose.

[0023] Allogeneic MSCs can be LOMECEL-B TM Cells, specifically Longeveron preparations of allogeneic human mesenchymal stem cells. Useful stem cells (including LOMECEL-B) TM Other uses and preparations of (branded mesenchymal cells) can be found in the following U.S. patent applications, all of which are incorporated herein by reference: US20190038742A1; US20190290698A1; and US20200129558A1. Attached Figure Description

[0024] Figure 1 This paper describes the consort study design, which summarizes the patient arrangement in a phase 2a double-blind, randomized, placebo-controlled 45-week trial to evaluate the effect of Lomecel-B on Alzheimer's disease.

[0025] Figure 2A The changes in CADS scores at week 0 (baseline), week 16, week 26, and week 39 were depicted in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0026] Figure 2B The changes in total MoCA scores at week 0 (baseline), week 16, week 26, and week 39 were depicted in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0027] Figure 2C The changes in total MMSE2 scores at week 0 (baseline), week 16, week 26, and week 39 were depicted in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0028] Figure 2D The changes in CDR-SB total score were depicted in four patient groups at weeks 0 (baseline), 4, 8, 12, 16, 26, and 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0029] Figure 2E The changes in the total ADAS-cog13 score were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0030] Figure 2F The changes in ADCS-ADL scores at weeks 0 (baseline), 16, 26, and 39 were depicted in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0031] Figure 2G The changes in QoL-AD (care provider) scores were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0032] Figure 2H The changes in QoL-AD (study subjects) scores in four patient groups at week 0 (baseline), week 16, week 26, and week 39 were depicted: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0033] Figure 2I The changes in ADRQL scores at weeks 0 (baseline), 16, 26, and 39 were depicted in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0034] Figure 3A MR images depicting changes in brain volume from patients in group 1 (placebo (dose × 4)) versus group 4 (MSC (100M × 4)) at week 39 were presented. During the 39-week trial period, the placebo group showed a steady decrease in total brain volume of approximately 1.2%, while the treatment group showed a statistically significant slowing of total brain atrophy.

[0035] Figure 3B The changes in whole brain volume at week 0 (baseline), week 16, week 26, and week 39 were depicted in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0036] Figure 3C The changes in gray matter volume (left) at week 0 (baseline), week 16, week 26, and week 39 were depicted in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0037] Figure 3D The changes in lateral ventricle volume (bilateral) were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0038] Figure 3E The changes in hippocampal volume (bilateral) at week 0 (baseline), week 16, week 26, and week 39 were depicted in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0039] Figure 3F The changes in temporal cortical volume (left) were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0040] Figure 3G The changes in medial temporal cortical volume (left) were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M ×4).

[0041] Figure 3H depicts the changes in mean diffusion rate (MD) in the bilateral cingulate cortex at weeks 0 (baseline), 16, 26, and 39 in four patient groups: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0042] Figure 4A The study demonstrated that improvements in bilateral hippocampal volume were significantly correlated with improvements in MMSE-2 scores.

[0043] Figure 4B The study demonstrated a significant correlation between improvements in whole-brain volume and improvements in MMSE-2 scores.

[0044] Figure 4C The study demonstrated that improvements in bilateral lateral ventricle volume were significantly associated with improvements in MMSE-2 scores.

[0045] Figure 4D The least-squares mean change in TIE2 from baseline was depicted in four patient groups at weeks 0 (baseline), 4, 8, 12, 16, 26, and 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0046] Figure 5A The changes in the thalamus (left) were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0047] Figure 5B The changes in the thalamus (right) were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0048] Figure 5C The changes in the frontal cortex (left) were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0049] Figure 5D The changes in the frontal cortex (right) were depicted in four patient groups at week 0 (baseline), week 16, week 26, and week 39: ● Placebo (dose × 4); MSC (25M × 1);■ MSC (25M × 4); MSC (100M × 4).

[0050] Figure 6A MR images of the cingulate cortex were depicted.

[0051] Figure 6B MR images of the medial temporal lobe cortex were depicted.

[0052] Figure 6C MR images of the cingulate cortex were depicted.

[0053] Figure 6D MR images of the medial temporal lobe cortex were depicted.

[0054] Figure 7A The study depicted a linear decrease in whole-brain contraction in the placebo group.

[0055] Figure 7B The study described a statistically significant improvement in Lomecel-B compared to placebo.

[0056] Figure 8A The volume of the lateral ventricle (left) is depicted, where an increase is associated with a decrease in brain volume.

[0057] Figure 8B The volume within the lateral ventricle (right) is depicted, with an increase correlated with a decrease in brain volume.

[0058] Figure 8C The mean changes in the left ventricle from baseline are depicted.

[0059] Figure 8D The mean changes in the right ventricle from baseline were depicted.

[0060] Figure 9AThe mean change in gray matter (left) ratio from baseline was depicted, showing a statistically significant improvement in the Lomecel-b group compared to placebo at week 39.

[0061] Figure 9B The average change in gray matter ratio (right) from the baseline is depicted.

[0062] Figure 10A The mean variation from baseline in the left medial temporal lobe cortex was depicted.

[0063] Figure 10A The mean variation from baseline in the right medial temporal lobe cortex was depicted.

[0064] Figure 11A Describe the changes in the volume measurement of the left gray matter.

[0065] Figure 11B The changes in the right gray matter volume measurement are depicted.

[0066] Figure 12A The changes in left medial temporal lobe cortical volume measurements were depicted, with the treatment arms showing numerical superiority at weeks 26 and 39.

[0067] Figure 12B The changes in right medial temporal lobe cortical volume measurements were depicted, with the treatment group showing numerical superiority at weeks 26 and 39.

[0068] Figure 13A Left hippocampal volume measurements were depicted, with all treatment groups showing numerical superiority at week 39.

[0069] Figure 13B The right hippocampal volume was measured, with all treatment groups showing numerical superiority at week 39.

[0070] Figure 13C The mean change in left hippocampal volume from baseline was depicted, with the treatment group showing numerical superiority at weeks 26 and 39.

[0071] Figure 13D The mean change in right hippocampal volume from baseline was depicted, with the treatment group showing numerical superiority at weeks 26 and 39.

[0072] Figure 14A The mean change in the left thalamus ratio from baseline was depicted.

[0073] Figure 14B The mean change in the right thalamus ratio from baseline was depicted.

[0074] Figure 14C The volume of the left thalamus was measured.

[0075] Figure 14D The volume of the right thalamus was measured.

[0076] Figure 15A The average diffusivity of white matter (both sides) was depicted.

[0077] Figure 15B The axial diffusivity of white matter (both sides) was described.

[0078] Figure 15C depicts the white matter (bilateral) FA.

[0079] Figure 15D depicts the white matter (bilateral) RD.

[0080] Figure 16A The mean diffusion rate of the cingulate cortex (bilateral) was described.

[0081] Figure 16B The axial diffusion rate of the cingulate cortex (bilateral) was described.

[0082] Figure 16C The cingulate cortex (bilateral) FA was described.

[0083] Figure 16D The cingulate cortex (bilateral) RD was described.

[0084] Figure 17A The average diffusion rate of the entire corpus callosum (both sides) was depicted.

[0085] Figure 17B The axial diffusion rate of the entire corpus callosum (both sides) was depicted.

[0086] Figure 17C The entire corpus callosum (bilateral) FA is depicted.

[0087] Figure 17D The entire corpus callosum (bilateral) RD is depicted.

[0088] Figure 18A The white matter (bilateral) FA was described.

[0089] Figure 18B The white matter (bilateral) RD was described.

[0090] Figure 19 The changes in cortical volume from the baseline were depicted.

[0091] Figure 20 The changes in low metabolic characteristics from baseline are described.

[0092] Figure 21 The changes in frontal cortex volume from baseline were depicted.

[0093] Figure 22 The changes in parietal cortical volume from the baseline were depicted.

[0094] Figure 23 The changes in temporal cortex volume (bilateral) from baseline were depicted.

[0095] Figure 24 The volume changes of the occipital cortex (bilateral) from the baseline were depicted.

[0096] Figure 25 The changes in cortical volume from the baseline cingulate cortex were depicted.

[0097] Figure 26 The changes in cortical volume of the medial temporal lobe from baseline were depicted.

[0098] Figure 27 The changes in thalamic (bilateral) volume from baseline were depicted.

[0099] Figure 28 The changes in striatal (bilateral) volume from the baseline were depicted.

[0100] Figure 29 The changes in the entire cortical arterial spin marker (ASL) from baseline were depicted.

[0101] Figure 30 The changes in spin markers (ASL) of the medial temporal lobe cortical arteries from baseline were depicted.

[0102] Figure 31 The changes in eosinophil chemokine-1 concentration from baseline data were depicted.

[0103] Figure 32 The changes in eosinophil chemokine-2 concentrations from baseline data were depicted.

[0104] Figure 33 Raw mean eosinophil chemokine-3 concentration data were depicted.

[0105] Figure 34 The changes in eosinophil chemokine-3 concentrations from baseline data were depicted.

[0106] Figure 35 The concentrations of eosinophil chemokine-3 in experimental groups 1–4 were depicted.

[0107] Figure 36 The changes in GIP (activity) from baseline data were depicted.

[0108] Figure 37 The GIP (activity) of experimental groups 1-4 was described.

[0109] Figure 38 The original average GIP concentration was depicted.

[0110] Figure 39 The changes in GIP concentration from baseline data are depicted.

[0111] Figure 40 The changes in GIPF concentration from baseline data are depicted.

[0112] Figure 41 The changes in TIE2 concentration from baseline data are depicted.

[0113] Figure 42 The TIE2 concentrations of experimental groups 1-4 were depicted. Detailed Implementation

[0114] Alzheimer's disease (AD) is currently incurable and progressive. The FDA recently (2021-2024) approved a small number of antibody therapies (Cummings, J., et al. BioDrugs 38, 5-22 (2024)) designed to treat AD by removing β-amyloid peptides (Aβ or AβP), including aducanumab, lecanemab, and donanemab. These antibodies have shown some efficacy in reducing cognitive and functional decline in patients with early-stage AD. Studies have found that aducanumab carries a 35% risk of Alzheimer's-related imaging abnormalities (ARIA), particularly ARIA edema (ARIA-E) and a 19% risk of ARIA hemorrhage (ARIA-H) (Doran, SJ & Sawyer, RP Front Neurosci 18, 1326784 (2024)) and was discontinued in 2024. Lemcanemab resulted in a 27% improvement in the Clinical Dementia Rating Scale sum of boxes (CDR-SB) at 18 months (Swanson, CJ, et al., Alzheimers Res Ther 13, 80 (2021)), but was associated with a 21.5% risk of ARIA (Honig, LS, et al. Alzheimers Dement (NY) 9, e12377 (2023)) and the potential for progression of brain atrophy (Alves, F., et al. Neurology 100, e2114-e2124 (2023); Cousin-Frankel, J. Science 380, 19 (2023)). Lemcanemab also resulted in a 26.4% infusion-related response rate in patients. Donemumab carries a 36.8% risk of ARIA, and drug-induced inflammation and cerebral hemorrhage have also been reported (Sims, JR, et al. JAMA 330, 512-527 (2023)). Beyond these limited examples, most AD treatments resolve symptoms but do not alter disease progression, partly because the development and onset of AD are not fully understood, and this uncertainty is the source of much controversy in the field. Indeed, while most researchers in the field of AD development and treatment acknowledge that AβP accumulation plays a role in disease progression, it remains unclear whether AβP accumulation is a cause of AD or simply a result of dysregulation of other cellular pathways due to aging.

[0115] Furthermore, although AβP accumulation and tau-mediated neurofibrillary tangle formation remain the defining pathological features of AD, contributions from non-amyloid proteins and non-tau have been identified in AD, which may lead to cerebrovascular deterioration and a strong neuroinflammatory component that contributes to neuronal death and brain atrophy (Schwab, C. & McGeer, PL J Alzheimers Dis 13, 359-369 (2008); Scheffer, S., et al. Arterioscler ThrombVasc Biol 41, 1265-1283 (2021)).

[0116] Brain atrophy in AD involves multiple brain regions and begins relatively early in disease progression, affecting extensive areas of the occipital, parietal, frontal, and temporal lobes, as well as the hippocampus, sometimes up to 8 years before AD diagnosis (Scahill, RI, et al. ProcNatl Acad Sci USA 99, 4703-4707 (2002); Traini, E., et al. J Alzheimers Dis76, 317-329 (2020); Apostolova, LG, et al. Arch Neurol 63, 693-699 (2006); Jia, J., et al. N Engl J Med 390, 712-722 (2024)). MRI imaging has revealed progressive atrophy at 39 weeks using volumetric MRI, affecting multiple brain regions and the entire brain volume, accompanied by an increase in small ventricular sizes (Figure 3). During disease progression, biomarkers of brain atrophy may include a reduction in volume of the whole brain, gray matter, temporal lobe cortex, medial temporal lobe cortex, hippocampus, frontal cortex, and / or thalamus, while an increase in volume may also be observed in the lateral ventricles, white matter, or cingulate cortex. Furthermore, mean diffusivity or free water measurements in the cingulate cortex may increase with disease progression. These biomarkers can be quantitatively assessed and evaluated over time using magnetic resonance imaging (MRI) during disease progression.

[0117] Another important biomarker associated with AD progression is soluble TIE2 (sTIE2). Tyrosine kinase (TIE2), possessing homologous domains to immunoglobulins and epidermal growth factor, is a homologous receptor for angiopoietin 1 and 2. It is expressed by endothelial cells, activates downstream signaling pathways promoting angiogenesis and anti-inflammation, and can be degraded into a soluble form (sTIE2) released into the bloodstream. Recent findings indicate that sTIE2 concentrations are increased in the serum of AD patients, suggesting that the shedding and subsequent inactivation of cell surface receptors reduces anti-inflammatory activity.

[0118] We were surprised to find that compositions containing allogeneic mesenchymal stem cells (MSCs) could combat the symptoms of Alzheimer's disease (AD). Treatment of subjects with AD symptoms with compositions containing allogeneic MSCs has been found to improve brain morphology and reduce brain atrophy, and to promote the expression of novel quantitative biomarkers (including, but not limited to, the previously mentioned MRI biomarkers and serum biomarkers) for the diagnosis and evaluation of AD progression and the effectiveness of treatments. These findings are surprising due to the ambiguity surrounding the pathogenesis of AD and the general practice in treating AD with allogeneic MSCs, which are typically reserved for those skilled in the art because they are not expected to perform well due to their inability to directly target β-amyloid and their short residence time in the human body. Their large size also leads to the assumption that they cannot cross the blood-brain barrier and reach sites of inflammation and injury.

[0119] Another advantage of using allogeneic MSCs in the treatment of AD is that they do not involve targeting a single pathway or biomarker, such as AβP accumulation. Instead, using allogeneic MSCs in AD treatment allows multiple pathways to be targeted at once, thereby halting or significantly slowing the progression of AD.

[0120] Based on the above surprising findings, one aspect of this application relates to a method for determining the efficacy of alleviating AD symptoms or treating Alzheimer's disease (AD) or inhibiting the progression of AD in subjects with symptoms of Alzheimer's disease (AD) by providing novel biomarkers, wherein one or more biomarkers in the subject are measured before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs).

[0121] In some implementations, the method includes determining the efficacy of alleviating symptoms of Alzheimer's disease (AD) in subjects, wherein one or more biomarkers in the subjects are measured before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs).

[0122] In some implementations, the method includes determining the efficacy of treating Alzheimer's disease (AD) in subjects with symptoms of AD, wherein one or more biomarkers in the subject are measured before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs).

[0123] In some implementations, the method includes determining the efficacy of inhibiting the progression of AD disease in subjects with symptoms of AD, wherein one or more biomarkers in the subjects are measured before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs).

[0124] In some implementations, the biomarker is an MRI biomarker.

[0125] In some embodiments, the method also includes measuring the cognitive function of subjects with AD symptoms before and after administration of a composition containing allogeneic MSCs.

[0126] In some embodiments, the method also includes assessing the quality of life of subjects with AD symptoms before and after administration of a composition containing allogeneic MSCs.

[0127] In some implementations, MRI biomarkers include volume measurements of the whole brain, lateral ventricles, gray matter, hippocampus, temporal cortex, medial temporal cortex, hippocampus, thalamus, white matter, cingulate cortex, frontal cortex, or combinations thereof.

[0128] In some embodiments, the method includes measuring the whole brain volume of a subject suffering from AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured whole brain volume. In a preferred embodiment, a decrease in the measured degree of whole brain volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in whole brain volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0129] In some embodiments, the method includes measuring the gray matter volume of a subject suffering from AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured gray matter volume. In a preferred embodiment, a decrease in the measured gray matter volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of gray matter volume reduction can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0130] In some embodiments, the method includes measuring the temporal cortex volume of a subject suffering from AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured temporal cortex volume. In a preferred embodiment, the degree of reduction in the measured temporal cortex volume after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in temporal cortex volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0131] In some embodiments, the method includes measuring the medial temporal cortex volume of a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured medial temporal cortex volume. In a preferred embodiment, the degree of reduction in the measured medial temporal cortex volume after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in medial temporal cortex volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0132] In some embodiments, the method includes measuring the hippocampal volume of a subject suffering from AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured hippocampal volume. In a preferred embodiment, a decrease in the measured hippocampal volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in hippocampal volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0133] In some embodiments, the method includes measuring the frontal cortex volume of a subject suffering from AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured frontal cortex volume. In a preferred embodiment, the degree of reduction in measured frontal cortex volume after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in frontal cortex volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0134] In some embodiments, the method includes measuring the thalamic volume of a subject suffering from AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured thalamic volume. In a preferred embodiment, a decrease in the measured thalamic volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in thalamic volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0135] In some embodiments, the method includes measuring the lateral ventricle volume of a subject suffering from AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is an increase in the measured lateral ventricle volume. In a preferred embodiment, a decrease in the degree of increase in measured lateral ventricle volume after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of decrease in lateral ventricle volume increase can be in the range of 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0136] In some embodiments, the method includes measuring the volume of white matter in a subject suffering from AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is an increase in the measured white matter volume. In a preferred embodiment, a decrease in the degree of increase in measured white matter volume after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indication of efficacy. The decrease in the degree of increase in white matter volume can be in the range of 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0137] In some embodiments, the method includes measuring the cingulate cortex volume of a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is an increase in the measured cingulate cortex volume. In a preferred embodiment, a decrease in the measured increase in cingulate cortex volume after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The decrease in the increase in cingulate cortex volume can be in the range of 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0138] In some embodiments, the method includes measuring the mean diffusion rate or free water content in the cingulate cortex of a subject with AD symptoms using diffusion tensor imaging (DTI) or a similar technique before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In a preferred embodiment, a decrease in the mean diffusion rate or free water content in the cingulate cortex via DTI or a similar technique after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy.

[0139] In other embodiments, the method includes measuring one or more whole blood, plasma, or serum biomarkers in subjects with AD symptoms before and after administration of a composition containing allogeneic MSCs.

[0140] In some embodiments, the serum or plasma biomarker comprises soluble TIE2 (sTIE2). In some embodiments, the biomarker is an increase in sTIE2 levels in serum or plasma. In a preferred embodiment, a decrease in sTIE2 levels in serum or plasma after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy.

[0141] In some embodiments, the composition comprising allogeneic mesenchymal stem cells (MSCs) used to relieve AD symptoms, treat AD, and / or inhibit the progression of AD disease may contain 20 × 10⁻⁶ MSCs. 6 One allogeneic MSC, 25 × 10 6 One allogeneic MSC, 100 × 10 6 One allogeneic MSC, or 20 × 10 6 Up to 100 × 10 6 One allogeneic MSC, preferably 25 × 10⁻⁶ 6 One MSC, or more preferably 100 × 10 6 MSC.

[0142] In some embodiments, a composition comprising allogeneic mesenchymal stem cells (MSCs) is administered to a subject via intravenous or intra-arterial infusion. The composition is used to alleviate symptoms of AD, treat AD, and / or inhibit the progression of AD.

[0143] In some implementations, a composition comprising allogeneic mesenchymal stem cells (MSCs) is administered monthly to alleviate AD symptoms, treat AD, and / or inhibit the progression of AD disease.

[0144] In some implementations, the method includes using one or more biomarkers to determine the efficacy of alleviating AD symptoms in subjects with symptoms of Alzheimer's disease (AD), including measuring the one or more biomarkers in the subjects before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs).

[0145] In some implementations, the method includes using one or more biomarkers to determine the efficacy of treating AD or inhibiting the progression of AD in subjects with symptoms of Alzheimer's disease (AD), including measuring the one or more biomarkers in the subjects before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs).

[0146] In some implementations, the method includes using one or more biomarkers, wherein the biomarker is an MRI biomarker.

[0147] In some embodiments, the method includes using one or more biomarkers to determine efficacy in alleviating AD symptoms, treating AD, or inhibiting the progression of AD in subjects with AD symptoms, and further wherein cognitive function in subjects with AD symptoms is measured before and after administration of a composition comprising allogeneic MSCs.

[0148] In some implementations, the method includes using one or more biomarkers to determine efficacy in alleviating AD symptoms, treating AD, or inhibiting the progression of AD in subjects with AD symptoms, and further, wherein the quality of life of subjects with AD symptoms is assessed before and after administration of a composition comprising allogeneic MSCs.

[0149] In some implementations, the method includes using MRI biomarkers, wherein the MRI biomarkers include volume measurements of the whole brain, lateral ventricles, gray matter, hippocampus, temporal cortex, medial temporal cortex, thalamus, white matter, cingulate cortex, frontal cortex, or combinations thereof.

[0150] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of the whole brain volume of a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured whole brain volume. In a preferred embodiment, a decrease in the measured degree of whole brain volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in whole brain volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0151] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of gray matter volume of a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured gray matter volume. In a preferred embodiment, a decrease in the measured gray matter volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indication of efficacy. The degree of gray matter volume reduction can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0152] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of temporal cortex volume in a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured temporal cortex volume. In a preferred embodiment, a decrease in the measured temporal cortex volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in temporal cortex volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0153] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of the volume of the medial temporal cortex of a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured volume of the medial temporal cortex. In a preferred embodiment, the degree of reduction in the measured volume of the medial temporal cortex after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in the volume of the medial temporal cortex can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0154] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of hippocampal volume in a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured hippocampal volume. In a preferred embodiment, a decrease in the measured hippocampal volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in hippocampal volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0155] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of the frontal cortex volume of a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured frontal cortex volume. In a preferred embodiment, a decrease in the measured frontal cortex volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of reduction in frontal cortex volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0156] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of thalamic volume in a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is a reduction in the measured thalamic volume. In a preferred embodiment, a decrease in the measured thalamic volume reduction after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of thalamic volume reduction can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0157] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of the volume of the lateral ventricles of a subject before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs) to a subject with AD symptoms. In some embodiments, the biomarker is an increase in the measured lateral ventricle volume. In a preferred embodiment, a reduced degree of decrease in the measured increase in lateral ventricle volume after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The degree of decrease in the increase in lateral ventricle volume can range from 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0158] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of white matter volume in a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is an increase in the measured white matter volume. In a preferred embodiment, a decrease in the degree of increase in the measured white matter volume after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indication of efficacy. The decrease in the degree of increase in white matter volume can be in the range of 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0159] In some embodiments, the method includes using MRI biomarkers, wherein the biomarkers include measurements of the volume of the cingulate cortex of a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In some embodiments, the biomarker is an increase in the measured volume of the cingulate cortex. In a preferred embodiment, a decrease in the degree of increase in the measured volume of the cingulate cortex after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy. The decrease in the degree of increase in the volume of the cingulate cortex can be in the range of 0% to 10%, 1% to 5%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than or equal to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%.

[0160] In some embodiments, the method includes using MRI biomarkers, wherein the MRI biomarkers include mean diffusion rate or free water measurements via diffusion tensor imaging (DTI) or similar techniques. In some embodiments, the mean diffusion rate or free water measurements are mean diffusion rate measurements in the cingulate cortex of a subject with AD symptoms before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs). In a preferred embodiment, a decrease in the mean diffusion rate or free water measurements in the cingulate cortex after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy.

[0161] In other embodiments, the method includes using one or more biomarkers, wherein the biomarkers are one or more of whole blood, plasma, or serum biomarkers. In some embodiments, the serum or plasma biomarker comprises soluble TIE2 (sTIE2). In some embodiments, the biomarker is an increase in sTIE2 levels in serum or plasma. In a preferred embodiment, a decrease in sTIE2 levels in serum or plasma after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs) is an indicator of efficacy.

[0162] In some embodiments, the method includes using one or more biomarkers, wherein a composition comprising allogeneic mesenchymal stem cells (MSCs) is administered to alleviate symptoms of AD, treat AD, and / or inhibit the progression of AD, and wherein said composition comprises 20 × 10 6 One allogeneic MSC, 25 × 10 6 One allogeneic MSC, 100 × 10 6 One allogeneic MSC, or 20 × 10 6 Up to 100 × 10 6 One allogeneic MSC, preferably 25 × 10⁶ 6 MSCs, and / or more preferably 100 × 10 6 MSC.

[0163] In some implementations, the method includes using one or more biomarkers, wherein a composition comprising allogeneic mesenchymal stem cells (MSCs) is administered to a subject via intravenous or intra-arterial infusion to alleviate symptoms of AD, treat AD, and / or inhibit the progression of AD disease.

[0164] In some implementations, the method includes using one or more biomarkers comprising a composition of allogeneic mesenchymal stem cells (MSCs) as one or more of a single dose, a monthly dose, and a repeated interval dose to alleviate symptoms of AD, treat AD, and / or inhibit the progression of AD disease.

[0165] Example

[0166] Example 1

[0167] The effects of Lomecel-B on Alzheimer's disease: a randomized, double-blind, placebo-controlled phase 2a trial.

[0168] Experimental Design: This double-blind, randomized, placebo-controlled 45-week trial (ClinicalTrials.gov:NCT05233774) enrolled patients (60-85 years old) with mild Alzheimer's disease (MMSE score 18-24); evidence of amyloid on positron emission tomography (PET) and brain MRI consistent with Alzheimer's disease.

[0169] Lomecel-B intravenous infusion was administered in four study groups: Group 1: placebo infusion, once a month for four months (N = 12); Group 2: 25 million cells on day 0, followed by three placebo infusions per month (25 million cells x 1; N = 12); Group 3: 25 million cells per month for four months (25 million cells x 4; N = 13); Group 4: 100 million cells per month for four months (100 million cells x 4; N = 11). Figure 1 ).

[0170] Demographic and baseline characteristics are summarized in Table 1.

[0171] Table 1: Demographic and baseline characteristics.

[0172]

[0173] Evaluate: To ensure safety, patients were monitored after each Lomecel-B infusion (e.g., on day 0 and week 4, week 8 and week 12).

[0174] To analyze the efficacy of treatment, the composite AD score (CADS) was calculated at baseline and at weeks 16, 26, and 39.

[0175] Neurocognitive, neuropsychiatric, QOL, ADL, and CSHA-CFS assessments were performed at screening, baseline, and at weeks 16, 26, and 39 to assess changes from baseline (CFB).

[0176] Biomarkers: Serum-based biomarkers associated with the potential pro-angiogenic and anti-inflammatory activity of Lomecel-B and Alzheimer's disease progression, including sTIE2, were measured at screening, baseline, day 0, and weeks 4, 8, 12, 16, 26, and 39. sTIE2 was analyzed using a Meso Scale Discovery (MSD) electrochemiluminescence (ECL) immunoassay analyzer (1300MESO QuickPlex SQ120) and an angiogenesis kit (Meso Scale Discovery; cat #K15190D-1). Quantification was performed using the MSD V-Plex assay, with duplicate samples taken at each patient visit.

[0177] Brain volume measurements were performed via MRI at screening and at weeks 16, 26, and 39 to assess changes in volume measurements of the hippocampus, whole brain size, ventricular volume, and other brain structures, each normalized for intracranial volume. Brain MRI was performed using a 3T (Tesla) scanner, with two imaging centers across 10 clinical centers, using one MRI scanner at each location throughout the trial.

[0178] Diffusion tensor imaging (DTI) via MRI was performed at screening and at weeks 16, 26, and 39 to assess changes in neuroinflammation.

[0179] Goals and endpoints: Primary objective: To demonstrate the safety of intravenous delivery of single or multiple doses of Lomecel-B in patients with mild AD.

[0180] Primary endpoint: The proportion of patients who experienced at least one treatment-related serious adverse event (TE-SAE) within four weeks after each infusion.

[0181] Secondary objective: To identify efficacy signals of single or multiple doses of Lomecel-B compared to placebo in a patient population.

[0182] Secondary endpoint: Change in composite AD score (CADS) from baseline to week 39, which also included z-scores of CDR-SB, ADAS-Cog-13, ADCS-ADL, and left hippocampal volume.

[0183] Statistical analysis: All statistical analyses were performed using SAS software (v.9.4). Safety analyses were conducted on the ITT population based on the actual treatment received. For the primary endpoint, Clopper-Pearson exact confidence intervals were calculated for the percentage of patients with any SAE within 4 weeks post-infusion in all treatment groups (Group 1 [placebo], Group 2 [25M × 1], Group 3 [25M × 4], and Group 4 [100M × 4]) and in the Lomecel-B pooled groups 2, 3, and 4 (25M × 1, 25M × 4, 100M × 4) and in the Lomecel-B pooled groups 3 and 4 (25M × 4, 100M × 4). Additional safety parameters (laboratory parameters, vital signs, ECG) were descriptively summarized by treatment group and assessment time.

[0184] The secondary endpoint CADS was calculated by combining z-scores of changes from baseline (to the study endpoint) across multiple assessments, including ADCS-ADL, CDR-SB, ADAS-Cog-13, and left hippocampal volume (normalized for intracranial volume). Each component included 25% of the final score (i.e., each equally weighted). Inverse measures of ADAS-Cog-13 and CDR-SB were used to match the directionality of the other measures regarding changes in improvement / decline. Statistical analysis of changes from baseline in CADS was performed using a pooled model for repeated measures (MMRM) analysis. The model included fixed effects of baseline values ​​for visit, treatment group (4-level variables), visit-by-treatment interaction, sex, and outcome parameters. Patients were included in the model as a random effect. At each visit, the least squares (LS) mean, standard error (SE), and 95% CI were obtained from the model for each treatment group (including the pooled Lomecel-B treatment group). At each visit, the mean LS difference, SE, 95% CI, and two-sided p-value were obtained for the differences between treatment groups relative to placebo (group 1) for each active treatment group (groups 2, 3, and 4). Pooled Lomecel-B treatment group effects relative to placebo were obtained from the same model. Because this study was a hypothesis-generating study, for CADS, a difference was considered statistically significant if the two-sided p-value was < 0.1, and further investigation was warranted. No adjustments were made for multiple comparisons, and no primary or secondary outcome data were excluded.

[0185] The efficacy was evaluated based on the results of the completed Phase 1 study of Lomecel-B (Brody, M., et al. AlzheimersDement 19, 261-273 (2023)). The efficacy of this study was insufficient to detect a statistically significant difference in CADS. This study was initially designed to detect a difference of 3.87 points in MMSE-2 between the Lomecel-B treatment groups and placebo at 39 weeks with 85% efficacy. During the study, following industry practice of using composite endpoints for AD, the key secondary endpoint was changed to CADS score. The efficacy of this study was insufficient to detect a statistically significant difference in CADS. However, assuming a two-sided α of 0.1, the efficacy for detecting a 50% slowdown in disease progression at 6 months was 36% based on the comparison of the three active groups (N = 36) versus placebo (N = 12). An effect size as small as 0.2–0.3 in CADS can be considered clinically significant. No interim analysis was performed.

[0186] The intention-to-treat (ITT) cohort includes all randomized patients who receive at least one full or partial dose of an IP (any infusion of Lomecel-B or placebo). The modified intention-to-treat (mITT) cohort includes all patients who are randomized and receive at least one full or partial dose of an IP (any infusion of Lomecel-B or placebo) and complete at least one post-baseline efficacy assessment (biomarker data or cognitive test).

[0187] Similar MMRM analyses were performed on exploratory efficacy endpoints for ADAS-cog-13, MMSE-2, ADCS-ADL, CDR-SB, MoCA, NPI, QOL-AD (both care providers and study subjects), ADCS-ADL, ADRQL, brain volume measurement (via MRI), and diffusion tensor imaging.

[0188] result: Main results This study met its primary endpoint (Table 2). The primary endpoint was assessed by measuring the proportion of patients experiencing at least one treatment-related serious adverse event (TE-SAE) within 4 weeks of each infusion. In addition, safety assessments included the incidence of all adverse events (AEs) and SAEs throughout the study, including changes from baseline in clinical laboratory measurements and physical examinations. Sequential brain MRI was performed and used to assess the occurrence of ARIAs. The lower confidence limit for each Lomecel-B group overlapped with the upper confidence limit for group 1 (placebo), indicating no statistically significant difference in TE-SAE rates between group 1 (placebo) and any Lomecel-B group within 4 weeks after infusion (95% CI for group 1: 0–26.5; groups 2 and 3: 0.2–36.0; and group 4: 0.2–41.3). Overall, the safety evaluation supports that Lomecel-B is safe and well-tolerated in the study population in both single-dose and multiple-dose regimens.

[0189] Table 2: Security Overview.

[0190]

[0191] Secondary results

[0192] The pre-specified secondary endpoint of this study was the change in the composite Alzheimer's disease score (CADS) from baseline (CFB) to week 39 in the Lomecel-B treatment group compared to placebo. CADS was calculated using the Alzheimer's Disease Collaborative Study Activity in Daily Life (ADCS-ADL), the Clinical Dementia Rating Scale Box Total Score (CDR-SB), the Alzheimer's Disease Assessment Scale-Cognitive Subscale 13 (ADAS-Cog-13), and a z-score of the left hippocampal volume via MRI. This is similar to other comprehensive scoring methods used to measure AD progression, including ADCOMS (Wang, J., et al. J Neurol Neurosurg Psychiatry 87, 993-999 (2016)), the Comprehensive AD Rating Scale (iADRS) (Wessels, AM, et al. J PrevAlzheimers Dis 2, 227-241 (2015)) and the Global Statistical Test (GST) (Huang, P., et al. MovDisord 24, 1732-1739 (2009)).

[0193] As depicted in Figure 2a, the significant progressive decrease in CADS scores in the placebo group compared to placebo was attenuated in group 2 (25M × 1) and in the combination groups consisting of patients from all active treatment groups (group 2: 25M × 1, group 3: 25M × 4, group 4: 100M × 4; hereinafter referred to as "combination treatment groups 2-4"), providing temporary support for slowing disease progression (change in group 2 compared to placebo: 0.38; 95% CI: -0.06–0.82; p = 0.091; pre-specified p < 0.1 as a positive result due to small sample size). When assessed using each regimen group, CADS values ​​were positive in group 2 (change: 0.39, 95% CI: -0.06–0.85, p = 0.086) and group 4 (change: 0.44, 95% CI: -0.07–0.95, p = 0.09) compared to placebo.

[0194] Exploratory results

[0195] All patients underwent a series of clinical assessments of cognition and function using the MoCA, MMSE-2, Clinical Dementia Rating Scale Box Total Score (CDR-SB), and the Alzheimer's Disease Assessment Scale-Cognitive Subscale 13 (ADAS-Cog-13) (Nasreddine, ZS, et al. J Am Geriatr Soc 53, 695-699 (2005); Cummings, JL, et al. Neurology 44, 2308-2314 (1994)). Compared with placebo, all Lomecel-B groups showed MoCA ( Figure 2B ) and MMSE-2 ( Figure 2C The trend of improvement in scores was statistically significant in MoCA (25M×1; p = 0.009; N = 12) in group 2 and in the combined treatment group (p = 0.015; N = 37). At week 39, there was a trend of improvement in MMSE-2 (100M ×4) in group 4 (p = 0.067; N = 11), but it did not affect CDR-SB or ADAS-Cog13 (Figures 2d and 2e). Nasreddine, ZS, et al. J Am Geriatr Soc 53, 695-699 (2005); Cummings, JL, et al. Neurology 44, 2308-2314 (1994).

[0196] Compared with the placebo group, ADCS-ADL scores showed improvement at week 39 in the high-dose group (group 4) and the combination therapy group (groups 2–4), with group 4 reaching statistical significance (100M × 4, p = 0.040; N = 11), and other Lomecel-B doses showed a trend towards improvement (Fig. 2f). Patients in group 4, assessed by care providers using QOL-AD, showed numerical improvement relative to group 1 (placebo) (Fig. 2g), but no improvement was observed in patient assessments (Fig. 2h). In ADRQL, no statistically significant difference was observed in the Lomecel-B treatment groups compared with placebo at week 39 (Fig. 2i).

[0197] MRI imaging, using volumetric MRI in the placebo group, revealed progressive brain atrophy at week 39, affecting multiple brain regions and the entire brain volume, accompanied by an increase in ventricular size (Fig. 3). Lomecel-B improved this decline relative to placebo in all treatment groups (Fig. 3). Representative vMRI images depict changes in brain volume at week 39 in group 1 versus group 4 (increase: yellow; decrease: blue) (Fig. 3a). Although the 25M × 1 treatment group (group 2) showed a non-statistically significant decrease in global brain atrophy, both the 25M × 4 and 100M × 4 groups (groups 3 and 4) showed statistically significant decreases in global brain atrophy at week 39 (group 3: P = 0.006; group 4: 0.009) (Fig. 3b). Compared to placebo, patients receiving Lomecel-B from combination therapy groups 2–4 showed a slower progression of global brain atrophy in 48% of cases (p = 0.005). At week 39, gray matter showed a numerical improvement relative to placebo (Fig. 3c). Ventricular enlargement, a characteristic feature of progressive AD decline, was observed in the placebo group (group 1). Compared with placebo, the Lomecel-B combination therapy group showed a numerical reduction of 34% in left ventricular enlargement (p = 0.097), and a numerical reduction of 37% in bilateral ventricular enlargement (p = 0.066) (Figure 3d). At week 39, the reduction in right ventricular enlargement in group 3 was statistically significant (N = 12; p = 0.042), and compared with placebo, the right ventricular enlargement in groups 2–4 was numerically reduced by 40% (N = 33; p = 0.044).

[0198] At week 39, bilateral hippocampal atrophy was significantly reduced in groups 2 and 3 (p = 0.029 and p = 0.028, respectively), while group 4 (p = 0.073) showed numerical improvement in bilateral hippocampal atrophy (Figure 3e). Combined Lomecel-B treatment in groups 2–4 also showed reductions in left, right, and bilateral hippocampal atrophy of 62% (p = 0.021), 53% (p = 0.073), and 59% (p = 0.013), respectively. In group 1 (placebo), the temporal cortex showed a significant decrease in volume, while treatment groups 2 and 4 showed a statistically significant slowing of this decrease compared to placebo (p = 0.028 and p = 0.042, respectively), and group 3 tended to improve (p = 0.076) (Figure 3f). The left medial temporal cortex showed progressive improvement toward week 39; although the reduction in atrophy in group 2 did not reach statistical significance, the high-dose groups 3 and 4 showed significant reductions in atrophy (group 3: p = 0.001; group 4: p = 0.032) (Figure 3g). Significant reductions in right medial temporal cortex atrophy were also observed at week 39 in groups 3 and the combination therapy group (p = 0.019).

[0199] The rate of atrophy in the parietal and occipital cortices was not significantly reduced. Similarly, no significant changes were observed in the cingulate cortex, striatum, and left thalamus compared to placebo, but the right thalamus showed a statistically significant reduction in atrophy (Group 4: P = 0.026). Figure 5A , Figure 5B By week 39, frontal cortical atrophy was significantly reduced in treatment group 4 (100M × 4) (left hemisphere: p = 0.028; right hemisphere: p = 0.002), but not in groups 2 and 3 (25M × 1 and 25M × 4; Fig. 5c, Fig. 5d).

[0200] Diffusion tensor imaging (DTI) has been used to indicate tissue structure and neuroinflammation and has become a valuable tool for assessing the neuropathology of Alzheimer's disease (Carlson, ML, et al. Alzheimers Dement (Amst) 13, e12218 (2021)). In this study, DTI (mean diffusion rate; MD) indicated that placebo in Alzheimer's disease may increase inflammation over 39 weeks compared to baseline (p = 0.011). In contrast, MD was reduced in all Lomecel-B treatment groups in the cingulate cortex compared to placebo, with group 2 reaching statistical significance (p = 0.048; Figure 3h). Sato, TN, et al. Nature 376, 70-74 (1995), Idowu, TO, et al. Elife 9 (2020).

[0201] Tyrosine kinase 2 (TIE2), possessing homologous domains to immunoglobulins and epidermal growth factor, is a homologous receptor to angiopoietin 1 and 2. It is expressed by endothelial cells, activates downstream signaling pathways promoting angiogenesis and anti-inflammation (Sato, TN, et al. Nature 376, 70-74 (1995)), and can be degraded into a soluble form (sTIE2) released into the bloodstream (Idowu, TO, et al. Elife 9 (2020)). The results of this study indicate that sTIE2 levels progressively increased in the placebo group compared to baseline (p = 0.047 at week 26), indicating the shedding and subsequent inactivation of cell surface receptors. Compared to placebo, Lomecel-B resulted in statistically significant reductions in serum sTIE2 levels in group 3 (25M × 4) at weeks 4 (p = 0.01), 8 (p = 0.02), and 16 (p = 0.045) (Figure 4d). sTIE2 levels also tended to decrease during treatment in groups 2 and 4. In all treatment groups, sTIE2 levels recovered towards placebo values ​​between weeks 26 and 39.

[0202] Post-event analysis

[0203] Pearson correlation analysis was performed to determine whether the reduced brain atrophy was associated with improved clinical scores. Improvements in bilateral hippocampal volume (R = 0.41; p = 0.008), global brain volume (R = 0.35; p = 0.023), and bilateral lateral ventricle volume (R = -0.35; p = 0.0213) were all significantly associated with improvements in MMSE-2 scores (Figures 4a-4c).

[0204] discuss

[0205] At week 16 (four weeks after final Lomecel-B treatment), statistically significant improvements in brain atrophy were not initially visible, but with subsequent follow-up, particularly at week 39, statistically significant changes became apparent. The reduction in brain atrophy was significant at the whole-brain level and enhanced in several brain regions. Regions showing the greatest improvement by week 39 included the temporal lobe and frontal lobe, with the temporal lobe encompassing the hippocampus, an area that demonstrates early atrophic progression and disease pathology in AD.

[0206] Lomecel-B is not expected to directly target β-amyloid or p-Tau, but rather the neurovascular and inflammatory components of AD and related conditions. Since this study did not report evidence of ARIA, Lomecel-B infusion could indicate a potentially new and safer treatment than anti-amyloid antibody therapy, which may be complementary and target a unique mechanism of the disease. Furthermore, Lomecel-B is presumed to express neurovascular protective and anti-inflammatory effectors, which have the potential to help counteract ARIA in combination therapy. Additionally, a strong correlation was observed between performance on cognitive scales and hippocampal volume retention. DTI imaging (mean diffusion rate) in the cingulate cortex, an area showing volume loss in early AD (Scheff, SW, et al. J Alzheimers Dis 43, 1073-1090 (2015); Planche, V., et al. Brain Commun 4, fcac109 (2022)), suggested a reduction in neuroinflammation, consistent with one of Lomecel-B's primary mechanisms of action. Previous evidence indicates that MSCs and their potential therapeutic effectors (e.g., exosomes) have activity for at least 7–10 days in animals and humans (Assis, AC, et al. Cell Transplant 19, 219–230 (2010); Preda, MB, et al. Cell Death Dis 12, 566 (2021); Kaushal, S., et al. EurHeart J Open 3, oead002 (2023); Gholamrezanezhad, A., et al. Nucl Med Biol 38, 961–967 (2011)), but in this study, the data suggest that there may be a more durable therapeutic benefit. Similar findings of sustained physiological / clinical improvement have been observed in studies of MSC administration to patients with congestive heart failure (Hare, JM, et al. JAMA 308, 2369-2379 (2012)) and age-related frailty (Tompkins, BA, et al. J Gerontol A Biol Sci Med Sci 72, 1513-1522 (2017)).

[0207] The results of this study indicate a significant increase in circulating TIE2 levels in the placebo group, which was offset by Lomecel-B. Consistent with previous studies, the decreased TIE2 levels observed in serum can reflect a reduction in the degradation and “shedding” of the extracellular portion of the TIE2 receptor following cleavage by metalloproteinases, which have been reported to be upregulated in AD (Idowu, TO, et al. Elife 9 (2020); Liao, MC & Van Nostrand, WEBiochemistry 49, 1127-1136 (2010); Yamada, T., et al. Acta Neuropathol 90, 421-424 (1995)). The increased TIE2 levels observed at week 26 support the role of TIE2 degradation in AD pathology. In principle, these findings are consistent with the pro-angiogenic / anti-inflammatory effects of Lomecel-B, mediated by circulating secretory factors that maintain TIE2 signaling in brain vascular endothelial cells (Joussen, AM, et al. Eye (Lond) 35, 1305-1316 (2021)), including but not limited to protein mediators and exosomes (Kaushal, S., et al. Eur Heart J Open 3, oead002 (2023); Behnke, J., et al. J Clin Med 9 (2020)). This has the potential to maintain BBB function and improve the brain's handling of amyloid and tau-related disease mechanisms. The time delay in observing rescue in local and global atrophy is consistent with the acute but long-lasting effects during treatment. Therefore, the effects of Lomecel-B may improve neurovascular unit function and alleviate neuroinflammation sufficiently to moderate the declining trajectory of neuronal death and atrophy during AD progression over time. It remains unknown whether the anti-inflammatory effect in AD is due to the direct anti-inflammatory effector produced by Lomecel-B, or indirectly due to the improvement in amyloid clearance through the maintenance of BBB function (Kinney, JW, et al. Alzheimers Dement (NY) 4, 575-590 (2018)).

[0208] Not all clinical and imaging biomarker results showed a clear dose-response relationship, but repeated dosing (groups 3 and 4) produced better responses than single dosing (group 1) in 27 of the 35 total clinical and vMRI measurements, and the highest-dose group (group 4) produced the best responses in 15 of those, indicating a bias towards escalating effect size with increasing dosing regimens. At the clinical level, care provider variability could explain some noise in the data, but on the other hand, volumetric MRI data can be considered more accurate. Besides small sample sizes, other factors contributing to data variability could include the variability in the potency of different batches of Lomecel-B, as potency may depend in part on the bone marrow donor.

[0209] By week 39, the areas showing the greatest improvement included the temporal lobe and the frontal lobe, with the temporal lobe including the hippocampus, which is an area that shows early atrophy progression and disease pathology in AD (Whitwell, JL Neurotox Res 18, 339-346 (2010)).

[0210] This study has limitations that need to be addressed. First, the sample size is relatively small, the majority of patients are Hispanic, and some variables, such as patients' education level, have not been determined. Second, the 39-week study duration is relatively short for a disease-modifying agent trial in an early-stage AD patient population, and longer studies are necessary. Furthermore, although the principles of global statistical testing support the use of composite endpoints in small AD studies, the CADS used here is not a validated composite endpoint and should be evaluated in future studies.

[0211] Research procedures and timelines.

[0212] Screening activities were conducted within 6 weeks prior to the first infusion therapy and included informed consent, medical history and physical examination collection, ECG, and clinical assessments (including MMSE-2 (Kueper, JK, et al. J Alzheimers Dis 63, 423-444 (2018)), ADAS-Cog-13 (Tzeng, RC, et al. Front Aging Neurosci 14, 1021792 (2022)), CDR-SB (Kahle-Wrobleski, K., et al. Alzheimers Dement (Amst) 6, 82-90 (2017)), QOL-AD (Potashman, M., et al. BMC Geriatr 23, 124 (2023)), and ADCS-ADL (Kasper, JD, et al. Alzheimer Dis Assoc Disord 23, 275-284). (2009)), ADRQL (Kasper, JD, et al. Alzheimer Dis Assoc Disord 23, 275-284 (2009)), NPI (Tompkins, BA, et al. J Gerontol A Biol Sci Med Sci 72, 1513-1522 (2017)), sample collection for safety laboratories, biomarker samples, and urine sample analysis. Screening procedures were performed using MRI and amyloid PET scan imaging (McKay, NS, et al. Nat Neurosci 26, 1449-1460 (2023)). Baseline visits were conducted within 4 weeks prior to the first infusion and included review of any changes in medical history, MoCA testing, reapplication of cognitive tests other than MMSE-2, and sample collection. Screening and baseline were spaced at least 2 weeks apart. All visit dates were relative to the first infusion schedule defined as time zero (0). Infusions were administered on day 0, week 4, week 8, and week 12. During infusion visits, 80 mL of Lomecel-B or placebo (80 mL of plasma lysate containing 1% HSA) was administered via peripheral intravenous infusion over 40 minutes. Prior to infusion, a review of accompanying medications and adverse events, as well as CDR-SB administration and sample collection, were performed. Follow-up visits were completed at weeks 16, 26, and 39, including clinical assessment and re-examination, ECG, patient and care provider assessment, sample collection, and MRI brain scan.

Claims

1. A method for determining the efficacy of alleviating symptoms of Alzheimer's disease (AD) in subjects, wherein one or more biomarkers in the subjects are measured before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs).

2. A method for determining the efficacy of treating AD or inhibiting the progression of AD in subjects with symptoms of Alzheimer's disease (AD), wherein one or more biomarkers in the subjects are measured before and after administration of a composition comprising allogeneic mesenchymal stem cells (MSCs).

3. The method according to claim 1 or 2, wherein the biomarker is an MRI biomarker.

4. The method according to any one of claims 1-3, wherein the method further comprises measuring the cognitive function or quality of life of the subject suffering from AD symptoms before and after administration of the composition comprising allogeneic MSCs.

5. The method according to claim 3, wherein the MRI biomarkers include volume measurements of the whole brain, lateral ventricles, gray matter, hippocampus, temporal cortex, medial temporal cortex, thalamus, white matter, cingulate cortex, frontal cortex, or combinations thereof.

6. The method of claim 5, wherein the volume measurement result is a measurement of the volume of the whole brain, gray matter, temporal cortex, medial temporal cortex, hippocampus, frontal cortex, or thalamus, and wherein the reduced degree of volume reduction measured after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indication of efficacy.

7. The method of claim 6, wherein after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs), the measured reduction in volume is reduced by 1% to 5%, 5% to 10%, 10% to 50%, or greater than 50%.

8. The method of claim 5, wherein the volume measurement result is a measurement result of the volume of the lateral ventricle, white matter, or cingulate cortex, and wherein a decrease in the degree of increase in volume measured after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indication of efficacy.

9. The method of claim 8, wherein after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs), the measured increase in volume is reduced by 1% to 5%, 5% to 10%, 10% to 50%, or greater than 50%.

10. The method according to any one of claims 3-9, wherein the MRI biomarker further includes average diffusion rate or free water measurement results by diffusion tensor imaging (DTI) or similar techniques.

11. The method of claim 10, wherein the average diffusion rate measurement is a measurement of the average diffusion rate in the cingulate cortex, and wherein a decrease in the average diffusion rate in the cingulate cortex after administration of the composition comprising allogeneic mesenchymal stem cells (MSCs) is an indication of efficacy.

12. The method of claim 1 or 2, wherein the method comprises measuring one or more whole blood, plasma, or serum biomarkers or multiple biomarkers in the subject suffering from AD symptoms before and after administration of the composition comprising allogeneic MSCs.

13. The method of claim 12, wherein the serum or plasma biomarker comprises soluble TIE2 (sTIE2), eosinophil chemokine 1, eosinophil chemokine 2, or eosinophil chemokine 3.

14. The method of claim 13, wherein a reduction in sTIE2 levels in the serum or plasma of the desired subject, measured after administration of the composition comprising allogeneic MSCs, is an indicator of efficacy.

15. The method according to any one of claims 1-14, wherein the composition comprises 25 × 10 6 MSC.

16. The method according to any one of claims 1-14, wherein the composition comprises 100 × 10 6 MSC.

17. The method according to any one of claims 1-16, wherein the composition is administered to the recipient via intravenous or intra-arterial infusion.

18. The method according to any one of claims 1-17, wherein the composition is administered to the recipient as one or more of a single dose, a monthly dose, and a repeat-interval dose.

Images