diff --git a/apis/python/src/tiledbsoma/io/ingest.py b/apis/python/src/tiledbsoma/io/ingest.py
index 7d87659a5a..28b34eac9b 100644
--- a/apis/python/src/tiledbsoma/io/ingest.py
+++ b/apis/python/src/tiledbsoma/io/ingest.py
@@ -13,6 +13,7 @@
 import time
 from typing import (
     Any,
+    ContextManager,
     List,
     Mapping,
     Optional,
@@ -24,6 +25,7 @@
     cast,
     overload,
 )
+from unittest import mock
 
 import anndata as ad
 import h5py
@@ -32,6 +34,7 @@
 import pyarrow as pa
 import scipy.sparse as sp
 import tiledb
+from anndata._core import file_backing
 from anndata._core.sparse_dataset import SparseDataset
 from somacore.options import PlatformConfig
 
@@ -104,6 +107,42 @@ def __init__(self, ingest_mode: str) -> None:
             )
 
 
+# This trick lets us ingest H5AD with "r" (backed mode) from S3 URIs.  While h5ad
+# supports any file-like object, AnnData specifically wants only an `os.PathLike`
+# object. The only thing it does with the PathLike is to use it to get the filename.
+class _FSPathWrapper:
+    """Tricks anndata into thinking a file-like object is an ``os.PathLike``.
+
+    While h5ad supports any file-like object, anndata specifically wants
+    an ``os.PathLike object``, which it uses *exclusively* to get the "filename"
+    of the opened file.
+
+    We need to provide ``__fspath__`` as a real class method, so simply
+    setting ``some_file_obj.__fspath__ = lambda: "some/path"`` won't work,
+    so here we just proxy all attributes except ``__fspath__``.
+    """
+
+    def __init__(self, obj: object, path: Path) -> None:
+        self._obj = obj
+        self._path = path
+
+    def __fspath__(self) -> Path:
+        return self._path
+
+    def __getattr__(self, name: str) -> object:
+        return getattr(self._obj, name)
+
+
+def _hack_patch_anndata() -> ContextManager[object]:
+    """Part Two of the ``_FSPathWrapper`` trick."""
+
+    @file_backing.AnnDataFileManager.filename.setter  # type: ignore
+    def filename(self: file_backing.AnnDataFileManager, filename: Path) -> None:
+        self._filename = filename
+
+    return mock.patch.object(file_backing.AnnDataFileManager, "filename", filename)
+
+
 # ----------------------------------------------------------------
 def from_h5ad(
     experiment_uri: str,
@@ -168,22 +207,21 @@ def from_h5ad(
     s = _util.get_start_stamp()
     logging.log_io(None, f"START  Experiment.from_h5ad {input_path}")
 
-    logging.log_io(None, f"START  READING {input_path}")
-
-    anndata = ad.read_h5ad(input_path, backed="r")
-
-    logging.log_io(None, _util.format_elapsed(s, f"FINISH READING {input_path}"))
-
-    uri = from_anndata(
-        experiment_uri,
-        anndata,
-        measurement_name,
-        context=context,
-        platform_config=platform_config,
-        ingest_mode=ingest_mode,
-        use_relative_uri=use_relative_uri,
-        X_kind=X_kind,
-    )
+    with tiledb.VFS(ctx=context.tiledb_ctx).open(input_path) as input_handle:
+        logging.log_io(None, f"START  READING {input_path}")
+        with _hack_patch_anndata():
+            anndata = ad.read_h5ad(_FSPathWrapper(input_handle, input_path), "r")
+        logging.log_io(None, _util.format_elapsed(s, f"FINISH READING {input_path}"))
+        uri = from_anndata(
+            experiment_uri,
+            anndata,
+            measurement_name,
+            context=context,
+            platform_config=platform_config,
+            ingest_mode=ingest_mode,
+            use_relative_uri=use_relative_uri,
+            X_kind=X_kind,
+        )
 
     logging.log_io(
         None, _util.format_elapsed(s, f"FINISH Experiment.from_h5ad {input_path} {uri}")
diff --git a/apis/python/tests/test_basic_anndata_io.py b/apis/python/tests/test_basic_anndata_io.py
index c882df5111..b0be11a784 100644
--- a/apis/python/tests/test_basic_anndata_io.py
+++ b/apis/python/tests/test_basic_anndata_io.py
@@ -219,16 +219,16 @@ def test_resume_mode(adata, resume_mode_h5ad_file):
     tempdir1 = tempfile.TemporaryDirectory()
     output_path1 = tempdir1.name
     tiledbsoma.io.from_h5ad(
-        output_path1, resume_mode_h5ad_file, "RNA", ingest_mode="write"
+        output_path1, resume_mode_h5ad_file.as_posix(), "RNA", ingest_mode="write"
     )
 
     tempdir2 = tempfile.TemporaryDirectory()
     output_path2 = tempdir2.name
     tiledbsoma.io.from_h5ad(
-        output_path2, resume_mode_h5ad_file, "RNA", ingest_mode="write"
+        output_path2, resume_mode_h5ad_file.as_posix(), "RNA", ingest_mode="write"
     )
     tiledbsoma.io.from_h5ad(
-        output_path2, resume_mode_h5ad_file, "RNA", ingest_mode="resume"
+        output_path2, resume_mode_h5ad_file.as_posix(), "RNA", ingest_mode="resume"
     )
 
     exp1 = _factory.open(output_path1)
@@ -274,7 +274,7 @@ def test_ingest_relative(h5ad_file_extended, use_relative_uri):
 
     tiledbsoma.io.from_h5ad(
         output_path,
-        h5ad_file_extended,
+        h5ad_file_extended.as_posix(),
         measurement_name="RNA",
         use_relative_uri=use_relative_uri,
     )
@@ -356,7 +356,7 @@ def test_ingest_uns_string_array(h5ad_file_uns_string_array):
 
     tiledbsoma.io.from_h5ad(
         output_path,
-        h5ad_file_uns_string_array,
+        h5ad_file_uns_string_array.as_posix(),
         measurement_name="RNA",
     )