Treatment of pathologies associated with down syndrome with Anti-FSH antibodies

Abstract

Disclosed is a method for treating for treating pathologies associated with Down syndrome (e.g., symptoms of Alzheimer's disease and metabolic abnormalities), preventing the onset of pathologies associated with Down syndrome in patients with Down syndrome, reducing cognitive or functional decline, or reducing symptom load in a subject having Down Syndrome, in need or at risk thereof, comprising administering to said subject a therapeutically effective amount of an anti-Follicle Stimulating Hormone (FSH) antibody, or an antigen-binding portion thereof.

Description

[0001] Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO

[0002] TREATMENT OF PATHOLOGIES ASSOCIATED WITH DOWN SYNDROME WITH ANTI-FSH ANTIBODIES

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Patent Application No. 63 / 709,054, filed October 18, 2024, the contents of which are incorporated herein by reference in their entirety.

[0005] STATEMENT REGARDING FEDERAL FUNDING

[0006] This invention was made with government support under U01 AG073148 awarded by the National Institutes of Health, National Institute on Aging. The government has certain rights in the invention.

[0007] TECHNICAL FIELD

[0008] The present disclosure is directed to the treatment or prevention of pathologies associated with Down syndrome. More particularly, the disclosure is directed to the treatment or prevention of symptoms and pathologies of Down syndrome with antifollicle stimulating hormone (FSH) antibody.

[0009] BACKGROUND

[0010] Down Syndrome (DS) is the most common chromosomal disorder in humans; roughly 1 in 700 babies are born with DS. People with DS have a partial or full extra copy of chromosome 21, resulting in an extra copy of up to 200 protein-coding genes (PCGs) and 400 additional regulatory transcripts. These genetic alterations result in enhanced vulnerability to many health problems, including bone frailty, obesity, visual impairments, thyroid dysfunction, and cognitive impairments. However, the most devastating health condition faced by DS patients may be Alzheimer’s Disease (AD), due to both the early onset of beta-amyloid accumulation, combined with the near certainty of developing AD dementia. These seemingly disparate problems are typically ascribed to dysregulation arising from many genes operating in parallel on different biological Attorney Docket No.: 27527-0229WO1

[0011] Client Ref No. : T-240709-WO systems. In clinical practice, this means diseases are treated as they arise, leading to increasing numbers of medications. This polypharmacy approach contributes to considerable clinical complexity for managing the co-occurring health conditions of DS4, and puts people with DS at elevated risk for drug interactions and side effects.

[0012] One notable biological difference observed in both males and females with DS is elevated levels of Follicle-Stimulating Hormone (FSH) from infancy through adulthood. FSH is best studied in the context of sexual development and fertility. In this context, the elevated FSH levels observed in people with DS is not viewed as a problem of clinical significance. However, our published work shows considerable FSH receptor (FSHR) expression in many tissues beyond the gonads, including. Substantially extending the biology of FSH, we established a causal link between normal, age-associated increases in FSH actions in brain and the genesis and progression of AD in mouse models. We also linked elevated FSH to bone frailty. Collectively, these observations suggest that a single biological change, early-onset elevated FSH, may underlie early-onset AD in DS, as well as multiple co-occurring disease vulnerabilities.

[0013] SUMMARY

[0014] Disclosed herein are methods for treating for treating pathologies associated with Down syndrome (e.g., symptoms of Alzheimer’s disease and metabolic abnormalities), preventing the onset of pathologies associated with Down syndrome in patients with Down syndrome, reducing cognitive or functional decline, or reducing symptom load in a subject having Down Syndrome, in need or at risk thereof, comprising administering to said subject a therapeutically effective amount of an anti -Follicle Stimulating Hormone (FSH) antibody, or an antigen-binding portion thereof.

[0015] In a first aspect, the disclosure provides methods for treating pathologies associated with Down syndrome, preventing the onset of pathologies associated with Down syndrome, reducing cognitive or functional decline in a subject with Down syndrome, or reducing symptom load of pathologies associated with Down syndrome in a subject in need or at risk thereof, comprising administering to the subject a therapeutically effective amount of an anti -Follicle Stimulating Hormone (FSH) antibody or an antigen-binding portion thereof, wherein the anti-FSH antibody, or antigen-binding Attorney Docket No.: 27527-0229WO1

[0016] Client Ref No. : T-240709-WO portion thereof comprises: (a) a heavy chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15; (b) a light chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 4, 12, 14, and 17; (c) a heavy chain CDR1 (CDRH1) comprising SEQ ID NO:5; (d) a heavy chain CDR2 (CDRH2) comprising SEQ ID NO:6; (e) a heavy chain CDR3 (CDRH3) comprising SEQ ID NO:7; (f) a light chain CDR1 (CDRL1) comprising SEQ ID NO:8; (g) a light chain CDR2 (CDRL2) comprising SEQ ID NO:9; and (h) a light chain CDR3 (CDRL3) comprising SEQ ID NO: 10. In some embodiments, the subject has a condition in which FSH levels are elevated. In some embodiments, the subject is female. In some embodiments, the subject is male. In some embodiments, the subject harbors trisomy 21 chromosomal abnormality and has been diagnosed with Down syndrome. In some embodiments, the pathologies associated with Down syndrome are selected from the group consisting of Alzheimer’s disease and metabolic abnormalities.

[0017] In some embodiments, the method alters one or more of the following in the subject in need thereof: (a) reduces Ap accumulation; (b) reduces amyloid plaques; (c) reduces Tau accumulation in the brain; and (d) enhances cognitive function.

[0018] In some embodiments, the one or more of Ap accumulation, amyloid plaques, and Tau accumulation in the brain is lower by at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, as compared to the corresponding reference levels in the subject or in a control.

[0019] In some embodiments, the cognitive function is enhanced by at least about 20%, at least about 30%, at least about 40%, or at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, as measured on one or more tests selected from the group consisting of the Alzheimer's Disease Assessment Scale- cognitive subscale (ADAS-cog); clinical global impression of change scale (CUBIC -plus scale); the Mini Mental State Exam (MMSE); the Neuropsychiatric Inventory (NPI); the Clinical Dementia Rating Scale (CDR); the Cambridge Neuropsychological Test Automated Battery (CANTAB); the Sandoz Clinical Assessment-Geriatric (SCAG), the Buschke Selective Reminding Test; the Verbal Paired Associates subtest; the Logical Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO

[0020] Memory subtest: the Visual Reproduction subtest of the Wechsler Memory Scale- Revised (WMS-R); the explicit 3- alternative forced choice task; and the Benton Visual Retention Test.

[0021] In some embodiments, the subject is concurrently treated with one or more agents selected from the group consisting of a cholinesterase inhibitor, an N-methyl-D-aspartate (NMD A) receptor antagonist, a hormone, a vitamin, an antipsychotic, a tricyclic antidepressant, a benzodiazepine, insulin, adeno-associated virus delivery of NGF, CERE- 110, beta-blocker, human amyloid vaccine, beta or gamma secretase inhibitor, nicotinic or muscarinic agonist, and a second antibody.

[0022] In some embodiments, the cholinesterase inhibitor is selected from the group consisting of galantamine, rivastigmine, tacrine, and donepezil. In some embodiments, the NMDA receptor antagonist is selected from the group consisting of ketamine, methadone, memantine, amantadine, and dextromethorphan or a salt thereof. In some embodiments, the antipsychotic agent is selected from the group consisting of aripiprazole, risperidone, olanzapine, quetiapine, or haloperidol. In some embodiments, the benzodiazepine is selected from the group consisting of lorazepam, oxazepam and temazepam. In some embodiments, the tricyclic antidepressant is nortriptyline. In some embodiments, the agent is a hormone selected from the group consisting of estrogen, progesterone and leuprolide. In some embodiments, the agent is a vitamin selected from the group consisting of folate and nicotinamide. In some embodiments, the second antibody is selected from the group consisting of bapineuzumab, solanezumab, gantenerumab, crenezumab, ponezumab, BAN2401, and aducanumab.

[0023] In some embodiments, the anti-FSH antibody or antigen-binding portion thereof is administered subcutaneously, intramuscularly, intravenously, intrathecally, or intracranially to the subject. In some embodiments, the anti-FSH antibody or antigenbinding portion thereof is administered to a subject in need thereof at a dose of about 0.2 to 50 mg / kg of the subject’s body weight. In some embodiments, the anti-FSH antibody or antigen-binding portion thereof is administered to a subject in need thereof twice a week, every week, every 2 weeks, every month, every two months, or every six months. In some embodiments, the anti-FSH antibody is administered continuously. Attorney Docket No.: 27527-0229WO1

[0024] Client Ref No. : T-240709-WO

[0025] In some embodiments, the cognitive decline is assessed by determining the subject’s score before and after administration of said anti-FSH antibody or antigenbinding fragment thereof, using an Alzheimer's Disease Assessment Scale-Cognition (ADAS- Cog) test. In some embodiments, the reduction in cognitive decline as measured by ADAS-Cog is at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to a placebo.

[0026] In some embodiments, the subject has mild, moderate, or severe AD. In some embodiments, the treatment is prophylactic for completely or partially preventing AD or symptoms thereof in the subject. In some embodiments, the treatment is therapeutic for partially or completely curing AD or symptoms associated with AD in the subject.

[0027] In another aspect, the disclosure provides pharmaceutical compositions comprising an isolated anti-Follicle Stimulating Hormone (FSH) antibody, or antigenbinding portion thereof, and a pharmaceutically acceptable carrier or excipient, wherein the composition is capable of crossing the blood brain barrier from the blood into the brain, wherein the anti-FSH antibody or antigen-binding portion thereof comprises: (a) a heavy chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15; (b) a light chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 4, 12, 14, and 17; (c) a heavy chain CDR1 (CDRH1) comprising SEQ ID NO: 5; (d) a heavy chain CDR2 (CDRH2) comprising SEQ ID NO: 6; (e) a heavy chain CDR3 (CDRH3) comprising SEQ ID NO: 7; (f) a light chain CDR1 (CDRL1) comprising SEQ ID NO:8; (g) a light chain CDR2 (CDRL2) comprising SEQ ID NO:9; and (h) a light chain CDR3 (CDRL3) comprising SEQ ID NO: 10.

[0028] In another aspect, the disclosure provides compositions for use in treating pathologies associated with Down syndrome, preventing the onset of AD in a subject with Down syndrome, or reducing cognitive or functional decline in a subject with Down syndrome, wherein the composition is administered intravenously, intrathecally, subcutaneously, or intracranially to the subject. In some embodiments, the composition is an isolated anti-Follicle Stimulating Hormone (FSH) antibody, or antigen-binding portion thereof, and a pharmaceutically acceptable carrier or excipient, wherein the composition is capable of crossing the blood brain barrier from the blood into the brain, wherein the Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO anti-FSH antibody or antigen-binding portion thereof comprises: (a) a heavy chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15; (b) a light chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 4, 12, 14, and 17; (c) a heavy chain CDR1 (CDRH1) comprising SEQ ID NO:5; (d) a heavy chain CDR2 (CDRH2) comprising SEQ ID NO:6; (e) a heavy chain CDR3 (CDRH3) comprising SEQ ID NO:7; (f) a light chain CDR1 (CDRL1) comprising SEQ ID NO:8; (g) a light chain CDR2 (CDRL2) comprising SEQ ID NON; and (h) a light chain CDR3 (CDRL3) comprising SEQ ID NO: 10.

[0029] In another aspect, the disclosure provides methods for treating for treating Alzheimer’s Disease (AD), preventing the onset of AD, reducing cognitive or functional decline, or reducing symptom load in a subject having Down Syndrome, in need or at risk thereof, comprising administering to said subject a therapeutically effective amount of an anti-Follicle Stimulating Hormone (FSH) antibody, or an antigen-binding portion thereof.

[0030] In another aspect, the disclosure provides methods for reducing FSH levels in a subject having Down Syndrome, comprising administering to said subject a therapeutically effective amount of an anti-Follicle Stimulating Hormone (FSH) antibody, or an antigen-binding portion thereof.

[0031] In another aspect, the disclosure provides methods for reducing beta-amyloid levels and improving cognitive performance in a subject having Down Syndrome, comprising administering to said subject a therapeutically effective amount of an anti- Follicle Stimulating Hormone (FSH) antibody, or an antigen-binding portion thereof.

[0032] In another aspect, the disclosure provides methods for treating obesity in a subject having Down Syndrome, comprising administering to said subject a therapeutically effective amount of an anti-Follicle Stimulating Hormone (FSH) antibody, or an antigenbinding portion thereof.

[0033] In another aspect, the disclosure provides methods for treating osteoporosis in a subject having Down Syndrome, comprising administering to said subject a therapeutically effective amount of an anti-Follicle Stimulating Hormone (FSH) antibody, or an antigen-binding portion thereof. Attorney Docket No.: 27527-0229WO1

[0034] Client Ref No. : T-240709-WO

[0035] In some embodiments, the anti-FSH antibody is Hu6. In some embodiments, the anti-FSH antibody or antigen-binding portion thereof is administered subcutaneously, intramuscularly, intravenously, intrathecally, or intracranially to the subject. In some embodiments, the anti-FSH antibody or antigen-binding portion thereof is administered to a subject in need thereof at a dose of about 0.2 to 50 mg / kg of the subject’s body weight. In some embodiments, the anti-FSH antibody or antigen-binding portion thereof is administered to a subject in need thereof twice a week, every week, every 2 weeks, every month, every two months, or every six months.

[0036] DESCRIPTION OF DRAWINGS

[0037] FIG. 1 shows a plot demonstrating that typical aging is associated with increases in FSH in both men and women.

[0038] FIG. 2 shows that FSH Acts on Hippocampal FSH Receptors (FSHRs) to Aggravate Alzheimer’s Disease (AD) in Mice. Expression by RT-PCR of FSHRs in human cortex, human neuro-blastoma cells (SH-SY5Y), rat cortical neurons, and mouse whole brain, cortex and hippocampus (A). Expression in mouse hippocampus, cortex and hypothalamus relative to ovary shown by qPCR (B), and FSHR protein in whole mouse brain by Western immunoblotting (C). Female 3xTg mice injected with FSH (5 IU daily, i.p. for 3 months) showed increased C / EBP0, AEP / 5-secretase, cleaved APP1'373and APP1-585, total Tau, cleaved Taul-368 and pTau in whole brain lysates (Western immunoblots, (D). This was consistent with cytokine release (E), and elevations in AEP activity (F), and Ap40 and A 42 (ELISA) in brain extracts (G). Electrophysiology for long-term potentiation showed reduced field excitatory post-synaptic potentials (fEPSPs) in the CAI hippocampal region in FSH-treated mice (H). Cognitive testing using the Morris Water Maze showed enhanced escape latency to mount the platform, increased integrated escape latency (AUC), and reduced percentage of time spent in the target quadrant (Probe Trial Test) (I). The findings were recapitulated in male 3xTg mice. In separate experiments, Fshr was knocked down in 3-month-old female 3xTg mice by stereotactic injection of AAV expressing short hairpin Fshr RNA or control shRNA, following which all mice were ovariectomized to induce AD. Western blots showed reduced C / EBPP, AEP, APP and Tau proteins, and FSHRs (J), findings that were in line Attorney Docket No.: 27527-0229WO1

[0039] Client Ref No. : T-240709-WO with improved cognition on Morris Water Maze (K). N-shown; *P<0.05, **P<0.01 . The compendium of biochemical, structural, functional, and electrophysiological data provide unequivocal evidence that FSH aggravates AD through a direct action on hippocampal FSHRs.

[0040] FIG. 3 shows that Anti-FSHp Antibody (FSH Ab) Alleviates AD in Two Mouse Models. We generated a polyclonal FSH Ab in goat against a 13-amino-acid mouse FSHp epitope, LVYKDPARPNTQK, that binds to the FSHR and blocks the action of FSH on FSHRs in bone and fat. To study the role of FSH as a driver of AD pathogenesis, 3xTg mice were ovariectomized (OVX) — this resulted in elevated serum FSH in the face of hypogonadism and induced AD pathology (A). OVX or sham-operated (Sham) mice were injected with FSH Ab or goat IgG (200 pg, i.p., every two days, for 8 weeks). FSH Ab reversed post-0 VX changes in C / EBPp, AEP, cleaved APP, Tau and phosphorylated Tau (B), Ap40 and Ap42 (ELISA) (C), AEP activation (D), apoptosis (TUNED; green) of hippocampal neurons (NeuN; red) (E), spine numbers in the CAI region (Golgi stained) (F), and hippocampal synapses (TEM, red arrows) (G). Cognitive testing by Morris Water Maze showed a reversal by the FSH Ab of altered escape latency, integrated escape latency (AUC), and percent time spent in the target quadrant (H). We replicated the 3xTg data using a second model — the APP / PS1 mouse. We used male instead of female mice and begun injecting FSH Ab at 5 months of age. Unlike 3xTg mice, APP / PS1 mice show a less pronounced phenotype and develop plaques at 6 months, and overt cognitive impairment at a later age of ~12 months. FSH Ab (120 to 150 pg, five times / week, i.p.) for 4 months reduced hippocampal and cortical A 4O and A042 levels (I). Mean ± SEM, N-shown, *P<0.05; **P<0.01. In all, the data establish that blocking FSH by an epitope-targeted Ab in hypogonadal female or eugonadal male AD mice prevents AD pathogenesis.

[0041] FIG. 4 shows the development of a humanized FSH blocking antibody. We started by generating two murine monoclonal anti-FSH0 antibodies, Mf4 and Hf2, against corresponding mouse and human FSHR-binding epitopes of FSH0, namely LVYKDPARPNTQK and LVYKDPARPKIQK; these were validated for FSH blocking activity. Hf2 with an IC50 of 6.1 nM in cell-based osteoclast assays was selected for humanization. Humanization: The variable domains of the heavy and light IgG chains, Attorney Docket No.: 27527-0229WO1

[0042] Client Ref No. : T-240709-WO

[0043] VH and VL, were amplified from Hf2 hybridomas. A mouse-human chimeric antibody, CHI, was first constructed by cloning the corresponding VH and VL, together with human IgGl-CH and IgK-CL fragments, respectively, into pTT5 vector. To generate fully humanized antibodies, a bacterial expression library consisting of the antigenbinding fragments (Fab) was produced with single site mutations introduced in the human framework flanking the complementarity-determining region (CDR), while keeping the CDR unaltered. This yielded 30 humanized Fab clones, which were expressed in E. coli. Crude supernatant extracts were initially tested for binding to both mouse and human FSH0 by ELISA, followed by confirmation of FSH binding by surface plasma resonance (SPR, Biacore) and rank-ordering by dissociation constants (Kds)15. Three full-length high-affinity humanized IgGs, Hu6, Hu26 and Hu28, with the lowest dissociation constants (Kds) were purified (A). We also digested Hu6 with papain to produce the Fab and Fc fragments that were purified using a protein A column (B). Initial Testing: Three complementary methods were used to document Hu6, Hu26 and Hu28 binding to human FSH SPR established Hu6 as having highest affinity for binding human FSH (lowest KD) (C) and was thus chosen as our lead candidate and purified to 98.97% (D). Binding was confirmed by ELISA (E), and the protein thermal shift assay (F). The latter used a fluorescent reporter Sypro-Orange to detect hydrophobic domains exposed following the unfolding of globular proteins. Hu6 showed a peak at ~69 °C due to Fc domain unfolding, and another at 77 °C from Fab unfolding (E). Incubation of Hu6 with human FSH produced a ~6 °C right-shift of the Fab peak, but not of the Fc peak. The Hu6-Fab fragment similarly showed a right-shift (3 °C) when incubated with FSH, with no shift with Hu6-Fc or LH. For atom-level detail, we modeled the variable regions of Hu6, Hu26 and Hu28 in complex with human FSH (Modeller 9.1); studied the antibody-FSHp interface (HADDOCK); identified interactions that stabilized the docked complexes (molecular dynamics); and calculated global electrostatic energy (AAG) (C). Interacting residues of human FSH are shown (G) — the Hu6: FSH model (H) shows interacting antibody (VH, VL) and FSHp residues. We studied the orientation of each residue of Hu6-Fab by estimating solvent accessible surface area (SAS A) (using GET AREA) (I). All interacting residues of Hu6 (arrows) were solvent accessible (SASA >20). We tested whether Hu6 blocked the interaction of FSH with the FSHR, for which we created a Attorney Docket No.: 27527-0229WO1

[0044] Client Ref No. : T-240709-WO stable FSHR-overexpressing HEK293 cell line. FSHR was detected by flow cytometry using Alexa647-labeled human FSH (*FSH) (J). Hu6, but not human IgG, reversed the right-shift arising from *FSH binding, indicating effective blockade. Consistent with this FSH-blocking action, Hu6 sharply attenuated the osteoclastogenic effect of FSH in a cell-based assay, whereas human IgG did not (K). A single i.p, injection of Hu6 into C57BL / 6J mice (N=6) yielded a Cmax of 20.5 pg / mL and t’A of -34.3 hours (in-house ELISA to detect human IgG in mouse serum) (L). No significant differences in serum LH, FSH activin or inhibin levels were noted over 4 days (D0-D4) after two consecutive FSH injections (M). Mean ± SEM, **P<0.01. Thus, our lead therapeutic, Hu6, binds human FSH with high affinity and blocks FSH action by preventing binding to the FSHR.

[0045] FIG. 5 shows data demonstrating that MS-Hu6 Distributes to Bone, Fat and Brain and other tissues of interest. 89Zr-MS-Hu6 was injected at 250 pCi (-250 pg) i.v. into C57BL / 6I mice followed by PET-CT scanning, which revealed persistence in multiple organs up to 72 hours (A). Maximal retention (SUVs, normalized to muscle) was noted in the liver, with persistence in regions of interest (ROIs), namely bone marrow, subcutaneous, and visceral white adipose tissue (WAT), and the brain (B). To confirm tissue persistence of89Zr-MS-Hu6, mice were perfused with PBS before tissue isolation for y-counting. Significant concentrations of89Zr-MS-Hu6 were detected in multiple organs, including bone, bone marrow, subcutaneous WAT (sWAT), visceral WAT (vWAT), brown adipose tissue (BAT) and brain (C). To study early events, we monitored the uptake by dynamic PET-CT imaging over 240 minutes. At 10 minutes, radioactivity was detected mainly in large vessels, followed at 60 and 240 minutes by organ permeation (D). Radioactivity was also detected in feces (E). To complement89Zr-based biodistribution studies, we labeled MS-Hu6 with Alexa-Fluor-750 (AF750), and injected C57BL / 6J mice. i.v. with AF750-MS-Hu6 (200 pg), AF750 alone or PBS. At 16 hours post-injection, mice were imaged using the IVIS platform. Significant soft tissue distribution of AF750-MS-Hu6 was noted (F). Upon perfusion and tissue isolation, there was AF750-MS-Hu6 uptake by the liver, kidney, fat depots, bone, and brain (G). In contrast, in the AF750 (dye only) control group, localization was noted only in the kidney due to dye excretion. In parallel, we also injected AF488-MS-Hu6 or unconjugated AF488 i.v. into C57BL / 6J mice (200 pg). Immunofluorescence was detected in the liver, Attorney Docket No.: 27527-0229WO1

[0046] Client Ref No. : T-240709-WO kidney, and hippocampal sections (H). Staining of hippocampal sections with an antihuman IgG confirmed brain localization (I). The biodistribution data in mice were replicated by injecting89Zr-MS-Hu6 i.v. into two Cynomolgus monkeys (J). The studies collectively demonstrate, using complementary technologies and two animal models, that injected MS-Hu6 localizes to and persists within bone, fat tissue and brain.

[0047] FIG. 6 shows that males in both the Ts65dn and Tc(HSA21,CAG-EGFP)l Yakaz / J mouse strains show elevated FSH (Fig. 6A; we have limited breeding colonies for this unfunded work and all trisomic females are used for breeding), consistent with other studies of Ts65Dn. Using male Ts65Dn mice at ~10 weeks of age, we measured FSH in plasma (A), fat mass with EchoMRI (B), bone mineral density with DXA (C), and glucose clearance with the glucose tolerance test (GTT). Some mice (n=3 / group) were implanted with mini-pumps which delivered either IgG or Hu6 (600pg per week delivered continuously) for 7 weeks total. The GTT was administered at Week 7 (D), and body weight was measured throughout (E). P-values reflect post-hoc comparisons (A-C), main effect of group (D), or a Group X Week interaction in a repeated-measures ANOVA (E). In the Ts65Dn line, we observe that trisomic males have increased adiposity (Fig. 6B), decreased BMD (Fig. 6C), impaired glucose clearance (Fig.6D) and continuous gain in body weight (Fig. 6E). When Hu6 was administered (s.c. via continuous infusion; 600pg / week), glucose clearance was greatly enhanced (Fig. 6D) and body weights remained stable throughout the experiment (Fig. 6E). These findings demonstrate our ability to use the implantable pumps in DS mice and also, though very underpowered, suggest that Hu6 can improve glucose clearance and reduce adiposity with only ~2 months of administration in young mice.

[0048] FIG. 7 shows plots of data demonstrating prevention and reversal of recognition memory loss by FSH blockade. NOR testing shows that H£2 prevents ovariectomy- induced memory loss in 3xTg mice (A), and restores lost memory in 22-month-old APP / PS1 mice (B).

[0049] FIG. 8 shows plots of data relating to body composition and bone mineral density (BMD), two measures sensitive to follicle-stimulating hormone (FSH) levels, in two mouse models of Down Syndrome. These data represent young (3-month) mice (A-C), and body composition and BMD in TcMAC21 mice (D-F). A) Young trisomic Ts65Dn Attorney Docket No.: 27527-0229WO1

[0050] Client Ref No. : T-240709-WO display greater fat mass than their wild-type (WT) littermates. Tn aged mice (8-months or greater), both groups of mice have much greater fat mass, and there is no longer a difference between WT and trisomic littermates. In contrast, both young (3-month) and older (5-month) trisomic Ts65Dn mice have lower BMD across the whole body (B) and tibia (C) compared to WT littermates. D) Older TcMAC mice have lower fat mass compared to their wild-type littermates. In contrast, older (5-month) trisomic TcMAC mice have lower whole body (E) and tibial (F) BMD compared to WT littermates.

[0051] FIG. 9 shows a plot of body weight in Ts65Dn and wild-type mice after administration of a humanized FSH-blocking antibody Hu6. Aged (>8-month) Ts65Dn or wild-type (WT) littermates were implanted with mini-pumps to deliver the humanized FSH-blocking antibody Hu6 (600pg / week) or IgG control continuously for 8 weeks. Delivery of Hu6 reduces body weight in the trisomic Ts65Dn mice, relative to IgG- treated WT littermates, who are gaining body weight over the same post-surgery period. These results suggest that even in older trisomic mice, who may have had longer exposures to elevated FSH than their WT littermates, an anti-FSH strategy provides health benefits by reducing FSH signaling.

[0052] Like reference symbols in the various drawings indicate like elements.

[0053] DETAILED DESCRIPTION

[0054] Unless otherwise defined herein, scientific and technical terms used in this application shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques of, pharmacology, cell and tissue culture, molecular biology, cell and cancer biology, neurobiology, neurochemistry, virology, immunology, microbiology, genetics and protein and nucleic acid chemistry, described herein, are those well-known and commonly used in the art. In case of conflict, the present specification, including definitions, will control.

[0055] The practice of the present application will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are within the skill of the art. Such techniques are explained fully in the literature, such as, Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989) Cold Spring Harbor Press; Attorney Docket No.: 27527-0229WO1

[0056] Client Ref No. : T-240709-WO

[0057] Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J.P. Mather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J.B. Griffiths, and D.G. Newell, eds., 1993-1998) J. Wiley and Sons; Methods in Enzymology (Academic Press, Inc.); Gene Transfer Vectors for Mammalian Cells (J.M. Miller and M.P. Calos, eds., 1987); Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Sambrook and Russell, Molecular Cloning: A Laboratory Manual, 3rd. ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2001); Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons, NY (2002); Harlow and Lane Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1998); Coligan et al., Short Protocols in Protein Science, John Wiley & Sons, NY (2003); Short Protocols in Molecular Biology (Wiley and Sons, 1999).

[0058] The nomenclatures used in connection with, and the laboratory procedures and techniques of biochemistry, immunology, microbiology, molecular biology, and virology described herein are those well-known and commonly used in the art.

[0059] Throughout this specification and embodiments, the word “comprise,” or variations such as “comprises or “comprising,” will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0060] It is understood that wherever embodiments are described herein with the language “comprising,” otherwise analogous embodiments described in terms of “consisting of’ and / or “consisting essentially of’ are also provided.

[0061] The term “including” is used to mean “including but not limited to.” “Including” and “including but not limited to” are used interchangeably.

[0062] Any example(s) following the term “e.g.” or “for example” is not meant to be exhaustive or limiting.

[0063] Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Attorney Docket No.: 27527-0229WO1

[0064] Client Ref No. : T-240709-WO

[0065] The articles “a,” “an,” and “the” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, an element means one element or more than one element. Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. For example, description referring to “about X” includes description of “X.” Numeric ranges are inclusive of the numbers defining the range. As used herein, the term “about” permits a variation of ±10% within the range of the significant digit.

[0066] Notwithstanding that the disclosed numerical ranges and parameters are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Moreover, all ranges disclosed herein are to be understood to encompass any and all subranges subsumed therein. For example, a stated range of “1 to 10” should be considered to include any and all subranges between (and inclusive of) the minimum value of 1 and the maximum value of 10; that is, all subranges beginning with a minimum value of 1 or more, e.g., 1 to 6.1, and ending with a maximum value of 10 or less, e.g., 5.5 to 10.

[0067] Where aspects or embodiments are described in terms of a Markush group or other grouping of alternatives, the present application encompasses not only the entire group listed as a whole, but each member of the group individually and all possible subgroups of the main group, and also the main group absent one or more of the group members. The present application also envisages the explicit exclusion of one or more of any of the group members in the Markush group or other grouping of alternatives.

[0068] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference in their entireties.

[0069] The term “about” refers to a range of values that would not be considered by a person of ordinary skill in the art as substantially different from the baseline values. For Attorney Docket No.: 27527-0229WO1

[0070] Client Ref No. : T-240709-WO example, the term “about” may refer to a value that is within 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value, as well as values intervening such stated values, for which context will define.

[0071] Exemplary methods and materials are described herein, although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the various aspects and embodiments. The materials, methods, and examples are illustrative only and not intended to be limiting.

[0072] Definitions

[0073] As used herein, the term “epitope” may refer to the region of an antigen to which an antibody or T cell binds, e.g. a region within the beta (P) subunit of FSH, including but not limited to an epitope within SEQ ID NO: 1 and / or SEQ ID NO: 2. An “antigen” refers to a substance that elicits an immunological reaction or binds to the products of that reaction.

[0074] As used herein, the terms “follicle stimulating hormone” and / or “FSH” may refer to a gonadotropin, a type of glycoprotein polypeptide hormone. FSH is synthesized and secreted by the gonadotropic cells of the anterior pituitary gland and is implicated in regulating the development, growth, maturation, and reproductive processes of the body. FSH is a 35.5 kDa glycoprotein heterodimer, having two polypeptide units, an alpha (a) and beta (P) subunit. FSH is similar in structure to luteinizing hormone (LH), thyroid- stimulating hormone (TSH), and human chorionic gonadotropin (hCG), sharing an identical alpha (a) subunit, but having variations in the beta (P) subunit. This makes the beta (P) subunit an attractive therapeutic target for FSH inhibitors as the inhibitors targeting the beta (P) subunit, e.g. one or more epitopes located within the beta (P) subunit, can be specific to inhibiting FSH. An exemplary gene encoding the beta (P) subunit of human FSH may be accessed at, e.g., Accession No. NM_000510. An exemplary gene encoding the beta (P) subunit of murine FSH may be accessed at, e.g., NM 008045. One of ordinary skill in the art will be able to reach predicted amino acid sequences from the provided nucleotide sequences.

[0075] As used herein, the term “FSH inactivating agent” is defined as an agent that reduces the bioactivity or bioavailability of FSH. The FSH inactivating agent may Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO achieve the reduction by binding directly or indirectly to FSH, such as in the case of anti- FSH antibodies or similar peptides or molecules, or the agent may prevent release of FSH from the anterior pituitary gland. In some embodiments, the FSH inactivating agent is a small molecule or a biologic (e.g., a soluble FSH receptor that can bind FSH and block its action). In the case of siRNAs, FSH inactivating agents may prevent translation of mRNA encoding FSH.

[0076] As used herein, the term “treating” or “treatment” of a disease may refer to executing a protocol, which may include administering one or more drugs to a patient (human or otherwise), in an effort to alleviate signs or symptoms of the disease. Thus, in the case of treating neurodegenerative diseases (such as Alzheimer’s disease), “treating” or “treatment” refers to taking steps to obtain beneficial or desired results, including clinical results, preventing the onset of the disease, reducing cognitive or functional decline in the disease, or enhancing cognitive function. Beneficial or desired clinical results include, but are not limited to, improving cognitive function, delaying or slowing the progression of cognitive impairment, reducing the rate of decline of cognitive function, preventing or slowing the progression of the disease or disorder, or alleviation, amelioration, or slowing the progression, of one or more symptoms associated with the disease. “Treating the disease also includes altering the levels of one of more pathogenic molecules in the disease, including but not limited to, reducing A accumulation; reducing amyloid plaques; and reducing Tau accumulation in the brain.

[0077] Down Syndrome

[0078] People with Down Syndrome (DS) have multiple hormonal abnormalities; amongst these is an elevated level of serum FSH in both males and females. This elevation of FSH has been observed very early in life, in both infants and young children. FSH levels are typically low in infancy, and increase at the time of puberty. In general, high levels of FSH are considered clinically benign, perhaps because increases of FSH are observed in women during the menstrual cycle and in both men and women as they age (Fig. 1). Elevated FSH can act on FSHRs in brain, adipocytes, and bone to induce AD pathology, obesity, and bone frailty, suggesting that long-term elevation of FSH, Attorney Docket No.: 27527-0229WO1

[0079] Client Ref No. : T-240709-WO even that observed in typical aging, can contribute to disease burden. People with DS have an increased prevalence of AD dementia, obesity, and bone frailty, which often emerges early in life. The lifetime risk of developing AD with DS is greater than 90%. Obesity rates are as high as 75% in adults with DS. Bone fracture prevalence is understudied in DS, but studies of those with intellectual disabilities (IDs) indicate the fracture rate in ID is roughly twice that of people without ID. The enhanced prevalence of AD, obesity and bone frailty at young ages in DS, combined with the observation that FSH is elevated in people with DS even in early ages, raises the intriguing possibility that FSH may be contributing to the co-occurrence of these health conditions in DS. The list of health conditions experienced by people with DS is substantial. We identified four additional health conditions that are shared across the three populations but understudied in mouse models of DS. These four conditions are: diabetes, thyroid dysfunction, impaired visual acuity, and pulmonary hypertension (see Table 1). Table 1. Health conditions in Down Syndrome and Turner Syndrome, two syndromes with elevated FSH. Attorney Docket No.: 27527-0229WO1

[0080] Client Ref No. : T-240709-WO

[0081] There are roles for FSH and FSHR in relevant tissues:

[0082] • Diabetes: FSHR has been detected in pancreas, and high levels of FSH were shown to drive insufficient insulin section in response to glucose. The incidence of diabetes in people DS is up to four times higher than that observed in control patients.

[0083] • Thyroid Dysfunction: FSHR has been detected in both follicular and parafollicular thyroid cells. Subclinical hypothyroidism, characterized by elevated thyroid stimulating hormone (TSH) and low levels of thyroxine (T4), is associated with elevated levels of FSH. People with DS have elevated rates of hypothyroidism, with a 39% prevalence in adults ages 18-29 and a 51% prevalence in those more than 30 years of age, whereas prevalence is estimated at ~5% across all adults in the US.

[0084] • Impaired Visual Acuity: FSHR is expressed in the cornea. At least 50% of people with DS have eye conditions that require glasses, including many that lead to decreased visual acuity and sensitivity. For example, keratoconus (KC) is a type of corneal disease that is estimated to impact somewhere between 5 and 71% of people with DS, but is very rare in people without DS. One study found elevated FSH levels in people with KC relative to people without KC, suggesting the possibility of a connection between elevated FSH and KC even outside of DS.

[0085] • Pulmonary Hypertension: FSHR is expressed at high levels in cardiomyocytes, endothelial cells of vessels around the heart, and the lungs. People with Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO

[0086] DS have elevated rates of pulmonary hypertension, with as many as 28% of children with DS experiencing the condition30 in addition to adults with DS.

[0087] Anti-FSH Antibodies and Antigen-Binding Fragments Thereof

[0088] The antibodies of the present disclosure are anti-FSH antibodies or antigenbinding fragments thereof. An anti-FSH antibody may take one of numerous forms in the art, as disclosed herein. Antibodies are in part defined by the antigens to which they bind, thus, an “anti-FSH antibody” is any such antibody which specifically binds at least one epitope found on FSH. In some embodiments, the epitope is located in the P subunit of FSH. In some embodiments, the epitope is located within LVYKDPARPKIQK (SEQ ID NO: 1). In some embodiments, the epitope is located within LVYKDPARPNTQK (SEQ ID NO: 2). It is understood in the art that an antibody is a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter - connected by disulfide bonds, or an antigen binding portion thereof. A heavy chain comprises a heavy chain variable region (VH) and a heavy chain constant region (CHI, CH2 and CH3). Alight chain comprises a light chain variable region (VL) and a light chain constant region (CL). The variable regions of both the heavy and light chains comprise framework regions (FWR) and complementarity determining regions (CDR). The four FWR regions are relatively conserved while CDR regions (CDR1, CDR2 and CDR3) represent hypervariable regions and are arranged from NH, terminus to the COOH terminus as follows: FWR1, CDR1, FWR2, CDR2, FWR3, CDR3, FWR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen while, depending of the isotype, the constant region (s) may mediate the binding of the immunoglobulin to host tissues or factors. It is known in the art that it is possible to manipulate monoclonal and other antibodies and use techniques of recombinant DNA technology to produce other antibodies or chimeric molecules which retain the specificity of the original antibody. Such techniques may evolve introducing DNA encoding the immunoglobulin variable region, or CDRs, of an antibody to the constant regions, or constant regions plus framework regions, of a different immunoglobulin. Attorney Docket No.: 27527-0229WO1

[0089] Client Ref No. : T-240709-WO

[0090] CDRs are defined by a variety of methods / systems by those skilled in the art. These systems and / or definitions have been developed and refined over a number of years and include Kabat, Chothia, IMGT, AbM, and Contact. The Kabat definition is based on sequence variability and generally is the most commonly used. The Chothia definition is based on the location of the structural loop regions. The IMGT system is based on sequence variability and location within the structure of the variable domain. The AbM definition is a compromise between Kabat and Chothia. The Contact definition is based on analyses of the available antibody crystal structures. An Exemplary system is a combination of Kabat and Chothia. Software programs (e.g., abYsis (www.bioinf.org.uk / abysis / sequence_input / key_annotation / key_annotation.cgi)) are available and known to those of skill in the art for analysis of antibody sequences and determination of CDRs. The specific CDR sequences defined herein are generally based on Kabat definitions. However, it will be understood that reference to a heavy chain CDR or CDRs and / or a light chain CDR or CDRs of a specific antibody will encompass all CDR definitions as known to those of skill in the art.

[0091] An exemplary anti-FSH antibody or antigen - binding portion thereof of the present disclosure comprises Hf2 or antigen binding portion thereof exemplified in US20190241651, incorporated by reference herein in its entirety. Hf2 has a variable heavy chain region comprising SEQ ID NO: 3 and a variable light chain region comprising SEQ ID NO: 4. Hf2 has a CDRH1 comprising SEQ ID NO: 5, a CDRH2 comprising SEQ ID NO: 6, a CDRH3 comprising SEQ ID NO: 7, a CDRL1 comprising SEQ ID NO: 8, a CDRL2 comprising SEQ ID NO: 9, and a CDRL3 comprising SEQ ID NO: 10. However, the anti-FSH antibodies or antigen-binding portions thereof of the present disclosure are not limited as such. For example, in some embodiments, the anti- FSH antibody has a variable heavy chain region having at least 70%, at least 75%, at least 80% , at least 85%, at least 90%, at least 95%, or at least 99% identity with SEQ ID NO: 3. In some embodiments, SEQ ID NO: 3 has at least one conservative substitution. In some embodiments, the anti-FSH antibody has a variable light chain region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with SEQ ID NO: 4. In some embodiments, SEQ ID NO: 4 has at least one conservative substitution. In some embodiments , one or more of the CDRs of the Attorney Docket No.: 27527-0229WO1

[0092] Client Ref No. : T-240709-WO variable heavy chain region has at least 70% , at least 75% , at least 80% , at least 85% , at least 90% , at least 95% , or at least 99% identity with one or more of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7. In some embodiments, one or more of SEO ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 has at least one conservative substitution. In some embodiments, one or more of the CDRs of the variable light chain region has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with one or more of SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10 . In some embodiments, one or more of SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10 has at least one conservative substitution.

[0093] Another exemplary anti-FSH antibody or antigen-binding portion thereof of the present disclosure comprises Hu6 or antigen binding portion thereof exemplified in Gera S. et al., PNAS, 117(46):28971-79. Hu6 has a variable heavy chain region comprising SEQ ID NO: 11 and a variable light chain region comprising SEQ ID NO: 12. Hu6 has a CDRH1 comprising SEQ ID NO: 5, a CDRH2 comprising SEQ ID NO: 6, a CDRH3 comprising SEQ ID NO: 7, a CDRL1 comprising SEQ ID NO: 8, a CDRL2 comprising SEQ ID NO: 9, and a CDRL3 comprising SEQ ID NO: 10. In some embodiments, the anti-FSH antibody has a variable heavy chain region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with SEQ ID NO: 11. In some embodiments, SEQ ID NO: 11 has at least one conservative substitution. In some embodiments, the anti-FSH antibody has a variable light chain region having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity with SEQ ID NO: 12. In some embodiments, SEQ ID NO: 12 has at least one conservative substitution.

[0094] Another exemplary anti-FSH antibody or antigen-binding portion thereof of the present disclosure comprises Hu26 or antigen binding portion thereof exemplified in Gera S. et al., PNAS, 117(46):28971-79. Hu26 has a variable heavy chain region comprising SEQ ID NO: 13 and a variable light chain region comprising SEQ ID NO: 14. Hu26 has a CDRH1 comprising SEQ ID NO: 5, a CDRH2 comprising SEQ ID NO: 6, a CDRH3 comprising SEQ ID NO: 7, a CDRL1 comprising SEQ ID NO: 8, a CDRL2 comprising SEQ ID NO: 9, and a CDRL3 comprising SEQ ID NO: 10. Yet another exemplary anti-FSH antibody or antigen-binding portion thereof of the present disclosure Attorney Docket No.: 27527-0229WO1

[0095] Client Ref No. : T-240709-WO comprises Hu28 or antigen binding portion thereof exemplified in Gera S. et al., PNAS, 117(46):28971-79. Hu28 has a variable heavy chain region comprising SEQ ID NO: 15 and a variable light chain region comprising SEQ ID NO: 16. Hu28 has a CDRH1 comprising SEQ ID NO: 5, a CDRH2 comprising SEQ ID NO: 6, a CDRH3 comprising SEQ ID NO: 7, a CDRL1 comprising SEQ ID NO: 8, a CDRL2 comprising SEQ ID NO: 9, and a CDRL3 comprising SEQ ID NO: 10.

[0096] The CDRs for the exemplified anti-FSH antibodies of this disclosure are as follows in Table 1:

[0097] Table 1: CDR sequences

[0098] The Variable Heavy (VH) and Variable Light (VL) chain sequences for the exemplified anti-FSH antibodies of this disclosure are as follows in Table 2:

[0099] Attorney Docket No.: 27527-0229WO1

[0100] Client Ref No. : T-240709-WO

[0101] Table 2: VH and VL sequences of the anti-FSH antibodies of this disclosure Attorney Docket No.: 27527-0229WO1

[0102] Client Ref No. : T-240709-WO

[0103] In some embodiments, the anti-FSH antibodies comprise the following CDR sequences shown in Table 3:

[0104] Table 3 : CDR sequences

[0105] The antibodies of the disclosure may comprise polyclonal antibodies. Methods of preparing polyclonal antibodies are known to the skilled artisan. Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent and, if desired, an adjuvant. Typically, the immunizing agent and / or adjuvant will be injected in the mammal by multiple subcutaneous or intraperitoneal injections. The immunizing agent may include the FSH polypeptide or a variant thereof. It may be useful to conjugate the immunizing agent to a protein known to be immunogenic in the Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO mammal being immunized. Examples of such immunogenic proteins include but are not limited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. Examples of adjuvants which may be employed include Freund's complete adjuvant and MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate). The immunization protocol may be selected by one skilled in the art without undue experimentation.

[0106] The antibodies may, alternatively, be monoclonal antibodies. Monoclonal antibodies may be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975). In a hybridoma method, a mouse, hamster, or other appropriate host animal, is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, the lymphocytes may be immunized in vitro.

[0107] The immunizing agent will typically include the FSH polypeptide or a variant thereof. The immunizing agent may comprise an antigenic fragment of the beta (P) subunit of FSH. The immunizing agent may be a peptide sequence comprising SEQ ID NO: 1 or SEQ ID NO: 2 or a peptide sequence consisting essentially of SEQ ID NO: 1 or SEQ ID NO:2 but having conservative substitutions. Generally, either peripheral blood lymphocytes (“PBLs”) are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice, Academic Press, (1986) pp. 59-103). Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine and human origin. Usually, rat or mouse myeloma cell lines are employed. The hybridoma cells may be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells. For example, if the parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (“HAT medium”), which substances prevent the growth of HGPRT-deficient cells. Attorney Docket No.: 27527-0229WO1

[0108] Client Ref No. : T-240709-WO

[0109] Preferred immortalized cell lines are those that fuse efficiently, support stable high level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, Calif, and the American Type Culture Collection, Manassas, Va. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, Marcel Dekker, Inc., New York, (1987) pp. 51-63). The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against FSH. Preferably, the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). Such techniques and assays are known in the art. The binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson and Pollard, Anal. Biochem., 107:220 (1980).

[0110] After the desired hybridoma cells are identified, the clones may be subcloned by limiting dilution procedures and grown by standard methods. Suitable culture media for this purpose include, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640 medium. Alternatively, the hybridoma cells may be grown in vivo as ascites in a mammal.

[0111] The monoclonal antibodies secreted by the subclones may be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

[0112] The monoclonal antibodies may also be made by recombinant DNA methods, such as those described in U.S. Pat. No. 4,816,567. DNA encoding the monoclonal antibodies of the disclosure can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells of the disclosure serve as a preferred source of such DNA. Once isolated, the DNA may Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. The DNA also may be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place of the homologous murine sequences, as in U.S. Pat. No. 4,816,567, or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide. Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the disclosure, or can be substituted for the variable domains of one antigen-combining site of an antibody of the disclosure to create a chimeric bivalent antibody.

[0113] The antibodies may be monovalent antibodies. Methods for preparing monovalent antibodies are well known in the art. For example, one method involves recombinant expression of immunoglobulin light chain and modified heavy chain. The heavy chain is truncated generally at any point in the Fc region so as to prevent heavy chain crosslinking. Alternatively, the relevant cysteine residues are substituted with another amino acid residue or are deleted so as to prevent crosslinking.

[0114] In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce fragments thereof, particularly, Fab fragments, can be accomplished using routine techniques known in the art.

[0115] In some embodiments, the antibodies or antigen binding fragments thereof of the present disclosure are covalently ligated to or enclosed within liposomes, albumin microspheres, microemulsions, nano-particles or nanocapsules. In some embodiments, the antibodies or antigen binding fragments thereof of the present disclosure are covalently ligated to a blood brain barrier protein, or conjugated to an antibody which binds to a receptor on blood vessels (such as an antibody to transferrin receptor (TfR), insulin receptor, leptin receptor, lipoprotein receptor, IGF receptor, LDL Receptor, P- selectin (CD62P), intracellular adhesion molecule- 1 (ICAM-1, or CD54), or other targets for cerebrovascular targeting), in order to facilitate blood-brain barrier penetration and delivery of the anti-FSH antibody or antigen binding fragment thereof to the disease site. Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO

[0116] Blood brain barrier shuttles, such as those disclosed in W02014033074 may be used to deliver the anti-FSH antibody or antigen binding fragment thereof to the brain.

[0117] Follicle-Stimulating Hormone

[0118] Follicle-stiumulating hormone (FSH) is elevated in Down Syndrome and a causative factor in Alzheimer’s pathogenesis, bone frailty, and adiposity. FSH is a pituitary hormone termed a gonadotropin — so named because it was originally characterized for its’ actions on testicles and ovaries. However, over the last two decades, our group has demonstrated important physiological roles for FSH outside of the gonads. FSH bypasses the gonads to act on bone directly. FSH regulates bone and elevated FSH is linked to bone frailty and osteoporosis. FSH also regulates body fat and energy expenditure. Typical aging is associated with increases in FSH in both men and women (Fig. 1), and FSH is associated with both bone mineral density (BMD) and body fat in humans. Fig 1. Data depicts the median ± 95% confidence intervals for FSH reference ranges from Siemen’s AD VIA Centaur FSH Assay.

[0119] FSH receptor (FSHR) is expressed throughout the brain. Elevated FSH is linked to AD in mouse models. Elevated FSH enhances the pathogenesis of AD (Fig. 2) and blocking the actions of FSH with a polyclonal anti-FSHp antibody prevents AD pathology and memory-related impairments (Fig 3).

[0120] FSH receptor (FSHR) is expressed extensively throughout the brain. Elevated FSH is linked to AD in mouse models. Elevated FSH enhances the pathogenesis of AD (Fig. 2) and blocking the actions of FSH with a polyclonal anti-FSHp antibody prevents AD pathology and memory-related impairments (Fig 3).

[0121] Pharmaceutical compositions

[0122] The anti-FSH antibodies of the present disclosure can be formulated as a pharmaceutical composition. Such pharmaceutical compositions can be formulated according to methods well known in the art; see, for example, Remington: The Science and Practice of Pharmacy (2000) by the University of Sciences in Philadelphia, ISBN 683-306472. The compositions can further comprise a pharmaceutically acceptable Attorney Docket No.: 27527-0229WO1

[0123] Client Ref No. : T-240709-WO carrier. Examples of suitable pharmaceutical carriers are well known in the art and include phosphate buffered saline solutions, water, emulsions, such as oil / water emulsions, various types of wetting agents, sterile solutions, etc. Additionally, the pharmaceutical compositions for injection may be prepared in lipophilic solvents, which include, but is not limited to, oils, such as vegetable oils, olive oil, peanut oil, palm oil soybean oil, safflower oil, etc.; synthetic fatty acid esters, such as ethyl oleate or triglycerides; cholesterol derivatives, including cholesterol oleate, cholesterol linoleate, cholesterol myristilate, etc.; or liposomes, as described above. The compositions may be prepared directly in the lipophilic solvent or preferably, as oil / water emulsions, (see for example, Liu, F. et al. Pharm. Res. 12: 1060-1064 (1995); Prankerd, R. J. J. Parent. Sci. Tech. 44: 139-49 (1990); U.S. Pat. No. 5,651,991).

[0124] In some embodiments, the anti-FSH antibodies of the present disclosure can be formulated as a pharmaceutical composition including, for example, one or more buffers, one or more salts, one or more surfactants, one or more sugars, and / or one or more lyoprotectants / cryoprotectants. The anti-FSH antibody formulation can include phosphate at a molar concentration of about 5 mM, about 10 mM, about 15 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, or about 50 mM. The anti-FSH antibody formulation can include Tween-20 at a concentration of about 0.001%, about 0.002%, about 0.003%, about 0.004%, or about 0.005% (v / v). The anti-FSH antibody formulation can include sodium chloride (NaCl) at a molar concentration of about 0.1 mM, about 0.2 mM, about 0.3 mM, about 0.4 mM, about 0.5 mM, about 0.6 mM, about 0.7 mM, about 0.8 mM, about 0.9 mM, about 1.0 mM, about 2.0 mM, about 3.0 mM, about 4.0 mM, or about 5.0 mM. The anti-FSH antibody formulation can include sucrose at a molar concentration of about 100 mM, about 110 mM, about 120 mM, about 130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, about 240 mM, about 250 mM, about 260 mM, about 270 mM, about 280 mM, about 290 mM, or about 300 mM.

[0125] The pharmaceutical composition may comprise additional agents. For example, for use in the treatment of pathologies associated with Down syndrome (e.g., symptoms of Alzheimer’s disease and metabolic abnormalities), the additional agent can be selected Attorney Docket No.: 27527-0229WO1

[0126] Client Ref No. : T-240709-WO from the group consisting of a cholinesterase inhibitor, an N-methyl-D-aspartate (NMD A) receptor antagonist, a hormone, a vitamin, an antipsychotic, a tricyclic antidepressant, a benzodiazepine, insulin, adeno-associated virus delivery of NGF, CERE- 110, beta-blocker, human amyloid vaccine, beta or gamma secretase inhibitor, nicotinic or muscarinic agonist, and a second antibody, such as an anti-Ap antibodies, anti-Tau antibodies, and combinations thereof.

[0127] The compositions of the disclosure can be administered through various routes known in the art, e.g., by intravenous, intraperitoneal, subcutaneous, intramuscular, topical, or intradermal administration.

[0128] The dose and dosage regimen depends upon a variety of factors readily determined by a physician, such as the severity of the condition, the patient, and the patient's history. Generally, a therapeutically effective amount of a composition is administered to a patient. In some embodiments, the amount of composition administered is in the range of about 0.1 mg / kg to about 100 mg / kg of patient body weight, and any range in between. Depending on the severity of condition, about 0.1 mg / kg to about 50 mg / kg body weight (for example, about 0.1-15 mg / kg / dose, more usually from about 0.2- 25 mg / kg body weight) of composition is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. The compositions may be delivered relatively low volume rates, for example but not necessarily from about 0.001 ml / day to 10 ml / day so as to minimize tissue disturbance or trauma near the site where the formulation is released. The formulation may be released at a rate of, depending on the specific biological agent(s), at a low dose, e.g., from about 0.01 pg / hr or 0.1 pg / hr, 0.25 pg / hr, 1 pg / hr, generally up to about 200 pg / hr, or the formulation is delivered at a low volume rate e.g., a volume rate of from about 0.001 ml / day to about 1 ml / day, for example, 0.01 micrograms per day up to about 20 milligrams per day. Dosage depends on a number of factors such as potency, bioavailability, and toxicity of the active ingredient and the requirements of the subject. The progress of this therapy is readily monitored by conventional methods and assays and based on criteria known to the physician or other persons of skill in the art. The above parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician. Attorney Docket No.: 27527-0229WO1

[0129] Client Ref No. : T-240709-WO

[0130] Methods of Treatment

[0131] The methods disclosed herein enable the treatment of pathologies associated with Down syndrome (e.g., symptoms of Alzheimer’s disease and metabolic abnormalities) in subjects with compositions of the disclosure. A composition of the disclosure comprises a recombinant, fully human, anti-FSH monoclonal antibody administered to the human patient. In a preferred embodiment, the monoclonal antibody has an excellent safety profde while being selective for FSH. A preferred monoclonal antibody meeting these criteria is the antibody comprising CDRs of SEQ ID NOs: 5-10. The antibody of the disclosure is a biologic treatment for pathologies associated with Down syndrome (e.g., symptoms of Alzheimer’s disease and metabolic abnormalities) which is a non-naturally occurring, recombinant, fully human, anti-FSH monoclonal antibody that recognizes FSH. A preferred antibody of the disclosure is an IgGl consisting of 2 heavy and 2 kappa light chains connected by inter-chain disulfide.

[0132] The methods of this disclosure further comprise administration of an anti-FSH antibody in combination with an additional therapeutic agent for a neurodegenerative disease (e.g., AD) or a metabolic syndrome. In some embodiments, an antibody of the disclosure is concurrently administered with one or more agents including but not limited to cholinesterase inhibitor, an N-methyl-D-aspartate (NMD A) receptor antagonist, a hormone, a vitamin, an antipsychotic, a tricyclic antidepressant, a benzodiazepine, insulin, adeno-associated virus delivery of NGF, CERE-110, beta-blocker, human amyloid vaccine, beta or gamma secretase inhibitor, nicotinic or muscarinic agonist, and a second antibody. The cholinesterase inhibitor includes but is not limited to galantamine, rivastigmine, tacrine, and donepezil. The NMDA receptor antagonist includes but is not limited to ketamine, methadone, memantine, amantadine, and dextromethorphan or a salt thereof. The antipsychotic agent includes but is not limited to aripiprazole, risperidone, olanzapine, quetiapine, or haloperidol. The benzodiazepine includes but is not limited to lorazepam, oxazepam and temazepam. The tricyclic antidepressant includes but is not limited to nortriptyline. The hormone includes but is not limited to estrogen, progesterone and leuprolide. The vitamin includes but is not limited to folate and nicotinamide. The second antibody includes but is not limited to Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO bapineuzumab, solanezumab, gantenerumab, crenezumab, ponezumab, BAN2401 , and aducanumab.

[0133] Furthermore, any therapy described herein can include one or more agents for treating, one or more side-effects of the therapy. Combination therapies (e.g., coadministration of an anti-FSH antibody composition and one or more additional neurodegenerative therapies or additional therapeutic agents) can be, e.g., simultaneous or successive. For example, the anti-FSH antibody of this disclosure and the additional therapeutic agent(s) can be administered at the same time or at different times. In some embodiments, the one or more additional therapeutic agents can be administered first in time and the anti-FSH antibody can be administered second in time.

[0134] Animal Models

[0135] Animal models serve as an important resource for developing and evaluating treatments for pathologies associated with Down syndrome (e.g., symptoms of Alzheimer’s disease and metabolic abnormalities). Features that characterize pathological hallmarks and cognitive impairment in animal models typically extend to pathological hallmarks cognitive impairment in humans (LaFerla FM, Green KN. Cold Spring Harb Perspect Med. 2012;2(l l):a006320. Published 2012 Nov 1.). Efficacy in such animal models is, thus, expected to be predictive of efficacy in humans.

[0136] In some embodiments, an animal model of pathologies associated with Down syndrome (e.g., symptoms of Alzheimer’s disease and metabolic abnormalities) can express one or more human genes. Animal models of Down syndrome can include the Ts65Dn or TcMAC21 mouse models. In some embodiments, an animal model can be the progeny of a wild-type (wt) animal and a transgenic (tg) animal. Such animal models can be used to assay the effectiveness of the methods and compositions of this disclosure in treating pathologies associated with Down syndrome (e.g., symptoms of Alzheimer’s disease and metabolic abnormalities).

[0137] In some embodiments, animal models may be used to evaluate the safety of a pharmaceutical composition, for example, to evaluate the safety of a pharmaceutical composition of an antibody therapy or combination therapy comprising antibodies and Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO one or more additional therapeutic agents. The use of animal models for evaluating the safety of a pharmaceutical composition can include investigating pharmacokinetics, pharmacodynamics, toxicology, efficacy, embryonic toxicity, carcinogenic potential, among other measures of safety of a pharmaceutical composition. Animal models for evaluating the safety of a pharmaceutical composition may include, for example, the use of mice, fish, frogs, rabbits, cats, dogs, or non-human primates.

[0138] We determine whether FSH is elevated in DS using the Ts65dn mouse line, which is trisomic for the human chromosome (Chr) 21 -homologous segment of mouse Chr 16 (corresponding to -70% of the trisomic genes in DS). We use the version of this strain missing a mutant gene that conferred retinal degeneration in the original strain. We also use the Tc(HSA21,CAG-EGFP)lYakaz / J mouse line, which is a model of human DS (-93% of PCGs and -79% of non-PCGs are triplicated). We find that males in both strains show elevated FSH (Fig. 6A), consistent with other studies of Ts65Dn7. Fig. 6: Using male Ts65Dn mice at -10 weeks of age, we measured FSH in plasma (A), fat mass with EchoMRI (B), bone mineral density with DXA (C), and glucose clearance with the glucose tolerance test (GTT). Some mice (n=3 / group) were implanted with mini-pumps which delivered either IgG or Hu6 (600pg per week delivered continuously) for 7 weeks total. The GTT was administered at Week 7 (D), and body weight was measured throughout (E). P-values reflect post-hoc comparisons (A-C), main effect of group (D), or a Group X Week interaction in a repeated-measures ANOVA (E).

[0139] In the Ts65Dn line, we observe that trisomic males have increased adiposity (Fig. 6B), decreased BMD (Fig. 6C), impaired glucose clearance (Fig.6D) and continuous gain in body weight (Fig. 6E). When Hu6 was administered (s.c. via continuous infusion; 600pg / week), glucose clearance was greatly enhanced (Fig. 6D) and body weights remained stable throughout the experiment (Fig. 6E). These findings demonstrate our ability to use the implantable pumps in DS mice and also suggest that Hu6 can improve glucose clearance and reduce adiposity with only -2 months of administration in young mice.

[0140] TcMAC21 mice are similar to Ts65Dn mice in that both show AD pathology and memory impairments. TcMAC21 mice show a decrease in fat mass, normal glucose Attorney Docket No.: 27527-0229WO1

[0141] Client Ref No. : T-240709-WO clearance, and lower body weight, primarily due to greater energy expenditure during the dark (awake) period.

[0142] EXAMPLES

[0143] The practice of the methods and compositions of the disclosure employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), cell culture, microbiology, cell biology, biochemistry, immunology, and neuroscience, which are well within the purview of the skilled artisan. Such techniques are explained fully in the literature, such as, "Molecular Cloning: A Laboratory Manual", second edition (Sambrook, 1989); "Oligonucleotide Synthesis" (Gait, 1984); "Animal Cell Culture" (Freshney, 1987); "Methods in Enzymology" "Handbook of Experimental Immunology" (Weir, 1996); "Gene Transfer Vectors for Mammalian Cells" (Miller and Calos, 1987); "Current Protocols in Molecular Biology" (Ausubel, 1987); "PCR: The Polymerase Chain Reaction", (Mullis, 1994); "Current Protocols in Immunology" (Coligan, 1991). These techniques are applicable to the methods and compositions of the disclosure. Particularly useful techniques for particular embodiments will be discussed in the sections that follow. The following materials, reagents, and methods are used for the Examples described herein.

[0144] Materials and Methods

[0145] AD Pathology

[0146] Hemisphere and cortex, cerebellar, and hippocampal sections are co-stained for Ap, TAU, pTAU, cleaved TAU and APP, NeuN, and TUNEL. All antibodies are available commercially. Galiyas Silver staining, thioflavin-S staining and Golgi staining for dendritic spines are carried out using standard protocols. ELISAs are performed for soluble Ap40 and A 42 isoforms (Invitrogen, Cat #s: KHB3481 and KHB3544).

[0147] Cognitive Performance

[0148] Novel Object Recognition (NOR) Test (long-term memory): A mouse is presented with two identical objects during the first session, and then one of the two Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO objects is replaced by a novel object during a second session. On day 1 , a habituation phase in an empty arena (5-min) is followed 24-hr later by training, which allows for a 5- min exploration in the habituated arena where two identical objects are placed in opposite quadrants. The testing phase, which follows a 20-min to 4-hr retention time, is followed by replacement of one object with a novel object and 5 to 10 min of exploration.

[0149] Parameters, including time spent, distance and number of head entries into quadrants, and object sniffing time, are collected using an ANY-Maze Video Tracking System (Stoelting, UK).

[0150] Nest building assessment (goal -directed attention): Mice are placed in a new cage containing a Nestlet for 4-hr and videotaped. The resulting nest quality will be scored according to 94. 3) Y maze test (working memory) Mice are placed in a Y maze with 34 x 8 x 14-cm arms and given 5 minutes to explore. The arms entered are measured. Because mice have the opportunity to do repeated entries into a single arm, there is a chance performance level of 22% (2 / 9) for spontaneous alternations.

[0151] Adiposity

[0152] Fat mass is measured with Quantitative Nuclear Magnetic Resonance. Live mice are placed in a thin-walled plastic cylinder, with freedom to turn around. An Echo3-in-l NMR analyzer (Echo Medical) was used to measure fat, lean and total mass, per manufacturer. Fat pads are also weighed at the time of sacrifice and body weights are taken at least once / week.

[0153] BMD

[0154] BMD is measured in live animals through DXA imaging, followed by pCT on post-mortem bone. Through an agreement between ISMMS and the U.S. Department of Agriculture (Grand Forks, North Dakota), we routinely collaborate with Jay Cao, PhD to perform pCT using Scanco VivaCT4. pCT parameters include BV / TV, Tb.Th, Tb.N, Tb.Sp, and Conn.D for trabecular bone (tibial metaphysis and spine); TMD-Cortical, Ct.BS, Ct.Pm, and Ct.Th for cortical bone (tibial diaphysis).

[0155] Glucose tolerance test (GTT) Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO

[0156] A glucometer is used to measure blood glucose in drops of tail blood. After baseline, mice are injected with 2g / kg b.w. of 20% glucose. Blood glucose is measured at 15, 30, 60, and 120-min post-injection.

[0157] Visual Acuity Sensitivity

[0158] Optomotor reflex (OMR) is measured. Mice are placed on a small platform in the middle of a box in which a rotating black and white striped pattern is presented at a constant velocity of 6.1° / s and maximal contrast. A camera installed above the platform automatically detects the mouse’s head movement in response to the presented stimulus. The software reduces the stripe width (to measure acuity) or the contrast between the strips (to measure sensitivity) and determines the thresholds at which the OMR is lost. All stimuli are presented and assessed binocularly.

[0159] Pulmonary function

[0160] Hemodynamic measurements (heart rate, cardiac output, right ventricle end- systolic pressure (RVSP) are recordede at the end of treatment. The lung and RV tissues are collected for histological and molecular profiling. The RV hypertrophy is calculated with the Fulton index (FI) and RV cardiomyocytes cross sections are measured. Pulmonary artery medial wall thickness is assessed by Hematoxylin and Eosin staining. The degree of vascular muscul arization is evaluated with aSMA antibody. In addition, apoptosis is assessed with TUNEL assay on lung sections.

[0161] Example 1: Determine the time course and mechanisms for elevated FSH in DS

[0162] Circulating FSH is measured at multiple early development time points after birth in a mouse model. Hormonal pathways that increase (activin B) and decrease (inhibin B, follistatin, estradiol, progesterone) FSH secretion from pituitary are measured. These indicate why FSH is elevated in DS when the Fsh gene is not one of the genes triplicated by DS. Attorney Docket No.: 27527-0229WO1 Client Ref No. : T-240709-WO

[0163] Example 2: Determine whether Hu6 prevents the development of Alzheimer’s pathology and impaired cognition, as well as and co-occurring metabolic and non- metabolic health conditions

[0164] Implantable, controllable osmotic mini pumps (Alzet) are used for long-term delivery of Hu6 or control IgG in young mice for either 2 or 4 months. Cognitive task performance, body composition, bone mineral density (BMD), visual acuity and sensitivity, thyroid function, and blood glucose clearance are measured both before and after treatment. At 6 months of age, upon treatment completion, AD pathology in multiple brain regions, bone density using Micro Computed Tomography (pCT), and pulmonary function are measured as terminal end points. These are all measures that depend on FSHR-expressing tissues and changes in these measures contribute to the co-occurring disease burden of DS.

[0165] Example 3: Determine whether Hu6 improves established Alzheimer’s pathology and impaired cognition, as well as co-occurring metabolic and non-metabolic health conditions

[0166] Hu6 or IgG are delivered to aged mice for either 2 or 4 months and the measures described in Example 2 are measured at 12 months of age.

[0167] Example 4: FSH Acts on Brain FSHR to Induce Pathology and Cognitive Decline

[0168] Gain- and loss-of-function experiments validating the target — FSH — were performed, demonstrating that FSH acts on brain FSHR to induce AD pathology and cognitive decline (Fig. 2). Fig. 3 provides proof-of-concept that an anti-FSH strategy rescues AD pathogenesis in both two models of AD. FSH inhibition also increases bone mass and reduces body fat, indicating that a single FSH-blocking agent can prevent and / or treat multiple disorders — AD, obesity, and osteoporosis. Fig. 4 shows the development of our humanized anti-FSH antibody, Hu6. Fig. 5 shows that Hu6 accesses multiple tissues where FSHR is strongly expressed, and that are tied to processes dysregulated in DS, including bone, adipose, brain, pancreas, and lung. Attorney Docket No.: 27527-0229WO1

[0169] Client Ref No. : T-240709-WO OTHER EMBODIMENTS

[0170] While the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. All references cited herein are incorporated by reference in their entireties.

Claims

FR RefNo.: 27527-0229WO1Client Ref No.: T-240709-WOCLAIMS1. A method for treating pathologies associated with Down syndrome, preventing the onset of pathologies associated with Down syndrome, reducing cognitive or functional decline in a subject with Down syndrome, or reducing symptom load of pathologies associated with Down syndrome in a subject in need or at risk thereof, comprising administering to the subject a therapeutically effective amount of an anti-Follicle Stimulating Hormone (FSH) antibody or an antigen-binding portion thereof, wherein the anti-FSH antibody, or antigen-binding portion thereof comprises:(a) a heavy chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15;(b) a light chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 4, 12, 14, and 17;(c) a heavy chain CDR1 (CDRH1) comprising SEQ ID NO:5;(d) a heavy chain CDR2 (CDRH2) comprising SEQ ID NO:6;(e) a heavy chain CDR3 (CDRH3) comprising SEQ ID NO:7;(!) a light chain CDR1 (CDRL1) comprising SEQ ID NO:8;(g) a light chain CDR2 (CDRL2) comprising SEQ ID NO:9; and(h) a light chain CDR3 (CDRL3) comprising SEQ ID NO: 10.

2. The method of claim 1, wherein the subject has a condition in which FSH levels are elevated.

3. The method of claim 1 or 2, wherein the subject is female.

4. The method of claim 1 or 2, wherein the subject is male.

5. The method of any one of claims 1-4, wherein the subject harbors trisomy 21 chromosomal abnormality and has been diagnosed with Down syndrome.FR RefNo.: 27527-0229WO1Client Ref No.: T-240709-WO6. The method of any one of claims 1-5, wherein the pathologies associated with Down syndrome are selected from the group consisting of Alzheimer’s disease and metabolic abnormalities.

7. The method of any one of claims 1-6, wherein the method alters one or more of the following in the subject in need thereof:(a) reduces Ap accumulation;(b) reduces amyloid plaques;(c) reduces Tau accumulation in the brain; and(d) enhances cognitive function.

8. The method of claim 7, wherein the one or more of Ap accumulation, amyloid plaques, and Tau accumulation in the brain is lower by at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, as compared to the corresponding reference levels in the subject or in a control.

9. The method of claim 8, wherein the cognitive function is enhanced by at least about 20%, at least about 30%, at least about 40%, or at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, as measured on one or more tests selected from the group consisting of the Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS- cog); clinical global impression of change scale (CIBIC-plus scale); the Mini Mental State Exam (MMSE); the Neuropsychiatric Inventory (NPI); the Clinical Dementia Rating Scale (CDR); the Cambridge Neuropsychological Test Automated Battery (CANTAB); the Sandoz Clinical Assessment-Geriatric (SC AG), the Buschke Selective Reminding Test; the Verbal Paired Associates subtest; the Logical Memory subtest: the Visual Reproduction subtest of the Wechsler Memory Scale- Revised (WMS-R); the explicit 3- alternative forced choice task; and the Benton Visual Retention Test.

10. The method of any one of the preceding claims, wherein the subject is concurrently treated with one or more agents selected from the group consisting of a cholinesterase inhibitor,FR RefNo.: 27527-0229WO1Client Ref No.: T-240709-WO an N-methyl-D-aspartate (NMD A) receptor antagonist, a hormone, a vitamin, an antipsychotic, a tricyclic antidepressant, a benzodiazepine, insulin, adeno-associated virus delivery ofNGF, CERE- 110, beta-blocker, human amyloid vaccine, beta or gamma secretase inhibitor, nicotinic or muscarinic agonist, and a second antibody.

11. The method of claim 10, wherein the cholinesterase inhibitor is selected from the group consisting of galantamine, rivastigmine, tacrine, and donepezil.

12. The method of claim 10, wherein the NMDA receptor antagonist is selected from the group consisting of ketamine, methadone, memantine, amantadine, and dextromethorphan or a salt thereof.

13. The method of claim 10, wherein the antipsychotic agent is selected from the group consisting of aripiprazole, risperidone, olanzapine, quetiapine, or haloperidol.

14. The method of claim 10, wherein the benzodiazepine is selected from the group consisting of lorazepam, oxazepam and temazepam.

15. The method of claim 10, wherein the tricyclic antidepressant is nortriptyline.

16. The method of claim 10, wherein the agent is a hormone selected from the group consisting of estrogen, progesterone and leuprolide.

17. The method of claim 10, wherein the agent is a vitamin selected from the group consisting of folate and nicotinamide.

18. The method of claim 10, wherein the second antibody is selected from the group consisting of bapineuzumab, solanezumab, gantenerumab, crenezumab, ponezumab, BAN2401, and aducanumab.FR RefNo.: 27527-0229WO1Client Ref No.: T-240709-WO19. The method of any one of the preceding claims, wherein the anti-FSH antibody or antigen-binding portion thereof is administered subcutaneously, intramuscularly, intravenously, intrathecally, or intracranially to the subject.

20. The method of any one of the preceding claims, wherein the anti-FSH antibody or antigen-binding portion thereof is administered to a subject in need thereof at a dose of about 0.2 to 50 mg / kg of the subject’s body weight.

21. The method of claim 20, wherein the anti-FSH antibody or antigen-binding portion thereof is administered to a subject in need thereof twice a week, every week, every 2 weeks, every month, every two months, or every six months.

22. The method of claim 20, wherein the anti-FSH antibody is administered continuously.

23. The method of any one of the preceding claims, wherein the cognitive decline is assessed by determining the subject’s score before and after administration of said anti-FSH antibody or antigen-binding fragment thereof, using an Alzheimer's Disease Assessment Scale-Cognition (ADAS- Cog) test.

24. The method of claim 23, wherein the reduction in cognitive decline as measured by ADAS-Cog is at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% relative to a placebo.

25. The method of any one of the preceding claims, wherein the subject has mild, moderate, or severe AD.

26. The method of any one of the preceding claims, wherein the treatment is prophylactic for completely or partially preventing AD or symptoms thereof in the subject.

27. The method of any one of the preceding claims, wherein the treatment is therapeutic for partially or completely curing AD or symptoms associated with AD in the subject.FR RefNo.: 27527-0229WO1Client Ref No.: T-240709-WO28. A pharmaceutical composition comprising an isolated anti-Follicle Stimulating Hormone (FSH) antibody, or antigen-binding portion thereof, and a pharmaceutically acceptable carrier or excipient, wherein the composition is capable of crossing the blood brain barrier from the blood into the brain, wherein the anti -FSH antibody or antigen-binding portion thereof comprises(a) a heavy chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 3, 11, 13, and 15 ;(b) a light chain variable sequence comprising a sequence that is at least 90% identical to a sequence selected from SEQ ID NOs: 4, 12, 14, and 17;(c) a heavy chain CDR1 (CDRH1) comprising SEQ ID NO:5;(d) a heavy chain CDR2 (CDRH2) comprising SEQ ID NO:6;(e) a heavy chain CDR3 (CDRH3) comprising SEQ ID NO:7;(f) a light chain CDR1 (CDRL1) comprising SEQ ID NO:8;(g) a light chain CDR2 (CDRL2) comprising SEQ ID NO:9; and(h) a light chain CDR3 (CDRL3) comprising SEQ ID NO: 10.

29. The composition of claim 26, for use in treating pathologies associated with Down syndrome, preventing the onset of AD in a subject with Down syndrome, or reducing cognitive or functional decline in a subject with Down syndrome, wherein the composition is administered intravenously, intrathecally, subcutaneously, or intracranially to the subject.

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